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Dynamic Light Scattering in Supramolecular

Materials Chemistry
J¨org Braun1 , Kasper Renggli1 , Julia Razumovitch1 ,
and Corinne Vebert2
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University of Basel, Basel, Switzerland
University of Geneva, Geneva, Switzerland

1 Introduction to Dynamic Light Scattering
2 Self-Processes
3 Supramolecular Reactivity and Devices
4 Soft Matter and Nanotechnology
5 Supramolecular Aspects of Chemical Biology
6 Conclusions
References

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INTRODUCTION TO DYNAMIC
LIGHT SCATTERING
Principle of dynamic light scattering

Light scattering (LS) is a powerful technique for characterizing the structure, structural formation, and interaction of
supramolecular systems (see Figure 1 for a schematic representation). Studying the scattering of light by structures
with sizes in the submicrometer range allows the determination of critical characteristics such as shape or internal
structure.
The absolute value of the intensity of the scattered
light is monitored in a static light scattering (SLS) mode,
whereas instantaneous variations in intensity are recorded
by dynamic light scattering (DLS). A combination of these

two LS modes yields complementary thermodynamic and
hydrodynamic information such as the molecular weight,
size, and shape of the system under investigation.1 In this
chapter, we focus on the use of DLS in supramolecular
materials science.
The detailed theory behind DLS, which is not the topic of
this review, is reported in several excellent publications.1–3
Briefly, electromagnetic waves—or photons—interact with
the local electron cloud of the analyte. This interaction
results in an energy transfer from the electromagnetic wave
to the electrons, inducing fluctuations, depending on the
polarizability of the analyte. Since any accelerated particle
emits light, the electrons reemit photons. This process,
extending from the interaction between the incident photon
to the reemission of light is called LS, and the analysis of
the characteristics of the scattered light yields information
about the system under investigation.
When a particle is subject to Brownian motion and
irradiated, two frequencies of equal intensity are generated
in addition to the frequency that would normally be
scattered, inducing a positive and a negative Doppler
shift proportional to the particle velocity. The interference
between the nonshifted wave (photon reemission) and the
two waves due to Brownian motion yields infinitesimal
variations in intensity. Detection of these is the basic
principle of DLS, which is therefore particularly suited to
the study of properties of solutions. The scattered intensity
is acquired as a function of time and is then self-correlated.
This yields the relaxation time due to the Brownian motion
and leads to the characterization of the particle size through
hydrodynamic models of the diffusion coefficients.

Supramolecular Chemistry: From Molecules to Nanomaterials, Online  2012 John Wiley & Sons, Ltd.
This article is  2012 John Wiley & Sons, Ltd.
This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons, Ltd.
DOI: 10.1002/9780470661345.smc039

Therefore. 1. Although these are polymeric systems. since they both allow the rapid and absolute determination of the diffusion coefficient. can only be accurately used for samples with low polydispersities. These include Taylor dispersion analysis (TDA). This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. in theory. on the other hand. and the determination of diffusion coefficients is then based on the assumption that systems with comparable sizes coelute.5 and size exclusion chromatography (SEC). it is possible to apply the findings to supramolecular ensembles. Ltd.2 Correlator Amplifierdiscriminator oto Ph iplier lt mu Computer storage Schematic representation of a typical light scattering Limitations to dynamic light scattering The intensity of the scattered light monitored over the course of an LS measurement depends on both the concentration and angle of detection. to accurately quantify the size of a particle. and is not disturbed by the presence of low-molecular-mass impurities. DLS is especially suited for the analysis of high-molecular-mass systems. the determination of the size of the molecules is hampered by DLS angle-dependent measurements. it is sometimes advisable to consider complementary techniques. the question arises as to whether an average diffusion coefficient still has any physical meaning. Therefore. a diffusion coefficient distribution should be given for polydisperse systems. However. DLS. is influenced much more by the polydispersity of the sample than TDA. The characterization of samples of low polydispersity was achieved best with TDA and DLS. Ltd. and SEC and showed that all four methods can be used effectively to determine diffusion coefficients of systems with low polydispersities by measuring a series of styrene acrylonitrile (SAN) copolymers. such as supramolecular systems. USAXS (ultra-small angle X-ray scattering) and USANS (ultra-small angle neutron scattering) can resolve Supramolecular Chemistry: From Molecules to Nanomaterials. HDC. However. As in cases of shape anisotropy. measurements must be carried out at several concentrations and angles. the scattered light from two different parts along the same molecule interferes constructively or destructively. Additional hydrodynamic methods include sedimentation velocity and viscosimetry. The most common model uses the Stokes–Einstein equation. the intensity of the scattered light does not exhibit angular dependence. here. 1. leading to an angle dependence of the intensity of the scattered light. The results of Mes indicate that this is indeed the case for SEC.1002/9780470661345. Both techniques are capable of achieving structural characterization of partially or fully ordered systems with sizes up to 150 nm and characteristic internal repeat distances between 5 and 25 nm. both of which provide information on the coarse structural properties of the system. these can be represented as single scattering centers.2 nm) compared with DLS since measurements are performed at very low angles (typically 0. spherical particles. The chromatographic methods of HDC and SEC have. Concentration-dependent measurements therefore allow extrapolation to infinite dilution for accurate size quantification by DLS.3 Alternatives to dynamic light scattering There are several other techniques complementary to DLS that allow the determination of the diffusion coefficient of supramolecular systems in dilute solution. HDC. however. might not always be the case.2 Techniques Cell Laser Bath − Figure 1 setup.7 Both methods require calibration with well-known standards. In the case of small particles S (S = laser wavelength/20. as the diffusion coefficient distribution for those samples spans an enormous range. The shape of the particle is assumed to be spherical. To fully characterize supramolecular systems. When particles cannot be described as point scattering centers. This article is  2012 John Wiley & Sons. DOI: 10. DLS. however. Mes and coworkers7 compared TDA.6 which can all be used if the supramolecular system has a low polydispersity. The use of small-angle X-ray scattering (SAXS)8 and smallangle neutron scattering (SANS)9 provides additional information on a much shorter wavelength scale (>0. in nanometer). Furthermore. Online  2012 John Wiley & Sons. and hydrodynamic models of the diffusion coefficient yield the equivalent hydrodynamic size of a sphere of the particle under investigation. rather than merely observing relative changes that depend on the composition of the surrounding medium. which assumes no intermolecular interactions between small. Ltd.1–10◦ ). Unlike TDA. for which the autocorrelation function must be corrected. When analyzing polydisperse samples. TDA has the disadvantage that it is subject to interference due to the presence of low-molecular-mass chromophoric compounds.smc039 . the use of DLS enables direct measurements of the Z-average diffusion coefficient of a polydisperse sample but requires a relatively large amount of the sample and is concentration dependent. however. few macromolecules are of a size that matches this size criterion.4 hydrodynamic chromatography (HDC). which. the ability to determine a relative diffusion coefficient distribution.

1002/9780470661345. DLS is a powerful tool for characterizing the properties of the ensemble. the size of the self-assembled particles increased. that is. the power law for the scattered light intensity indicates that the aggregates are fractal objects. therefore. Interestingly.24–26 However.13–18 Aside from linear copolymers. In several systems. local structural changes can be monitored by “point probes. At the shorter wavelength regime accessible to SAXS. a plethora of self-assembling copolymers can be synthesized according to various chemistry routes. drugs.1 K (1/A) Figure 2 Graphical representation of the scattered light intensity versus the scattering angle.2 :6 . This article is  2012 John Wiley & Sons. and FTIR (Fourier transform infrared) spectroscopy. Combining DLS. depend on several macromolecular properties such as molecular weight. and hydrophobic to hydrophilic volume fraction. Ltd. a delicate balance between steric. they were able to show that if the particles are loaded with small molecules. the micelle size decreases. The sizes of the core–shell-type nanoparticles.13 and several excellent review publications describe the self-assembly of block copolymers. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. for example. held together by noncovalent interactions. In this context. Schaefer and coworkers10 investigated the combined use of DLS and SAXS for the characterization of colloidal silica. attractive. DOI: 10. using poly(styrene) and poly(ethylene) polymer segments— either covalently linked (PS-b-PEO) or held by a bis(2. and repulsive forces drives the organization of the molecules into nanosized objects of various shapes such as vesicles or spherical. such as polar/hydrophobic monomers or rigid/flexible polymer segments. Online  2012 John Wiley & Sons. The characteristics of the resulting self-assembly. 1 H NMR (proton nuclear magnetic resonance). 19–21 Mai et al. intermolecular interactions were. These are composed of chemically or physically incompatible units along the same macromolecule. Ltd. An alternative to the connection between the polymer segments via metal ions to achieve an effective noncovalent self-assembly is molecular recognition among specially designed ligands and receptors. the pH or solvent composition itself influences the properties of self-organized structures by affecting the intermolecular interactions between the molecules building those systems.and rodlike core–shell micelles.23 produced self-processed nanoparticles from poly(ε-caprolactone) (PCL) grafted poly(vinyl alcohol) (PVA) copolymer to immobilize both hydrophobic and hydrophilic molecules. such as size or morphology.13. By combining both the techniques. micelles. The chemical incompatibility between covalently linked hydrophobic and hydrophilic polymer segments drives the organization of the macromolecules into 3 supramolecular structures. Self-processes are inherent in the self-assembly of copolymers. In Figure 2. Further. even larger dimensions.27–29 demonstrated that covalent binding between the hydrophobic and hydrophilic blocks is not mandatory to selfassemble copolymer structures.smc039 . The most widely investigated self-assembling copolymers are amphiphilic linear block copolymers.1 Intensity Light SAXS a−1 0. Since the resulting supramolecular assemblies are composed of several molecules. polydispersity. As the core-to-arm ratio increases. However. respectively.01 −4. Ltd. DLS in combination with transmission electron microscopy (TEM) enabled the determination of PEHOstar-PPO aggregates that self-assembled into large spherical micelles of defined sizes. few conventional techniques can be used to analyze their properties. sophisticated copolymer macromolecular architectures are also achievable. 16. large supramolecular aggregates can be fully characterized through the determination of the properties of the ensemble (DLS) and internal structure (SANS or SAXS). since far more energy and higher safety arrangements are needed.0 0. The disadvantage compared to DLS is intrinsic.12 2 SELF-PROCESSES Self-processes trigger the spontaneous organization of molecules into a higher ordered structure. the power law indicates that the monomer units that make up the aggregate remain intact. they showed the presence of large compound micelles resulting from the aggregation of single micelles interacting through hydrogen bonds. In dilute aqueous solution.22 synthesized hyperbranched multiarm copolyethers of 3ethyl-3-(hydroxymethyl)oxetane-propylene oxide (PEHOstar-PPO) of different molar ratios of PPO arms to PEHO cores. Owing to the constant progress in polymer chemistry.0001 0. were analyzed by DLS and TEM. Gohy et al. The Supramolecular Chemistry: From Molecules to Nanomaterials. frozen by covalent binding of the self-assembled molecules. Sheikh et al.001 0.” using methods such as circular dichroism (CD)11 and fluorescence techniques. They showed.2 -terpyridine)ruthenium(II) complex (PS-[Ru]PEO)—that both resulting copolymers self-assemble into similar morphologies.DLS in supramolecular materials chemistry −2.

the peptide self-assembled into discrete and stiff nanorods.smc039 . The shrinkage of polymer networks held by supramolecular interactions was controlled owing to the dual-complexing ability of the cyclodextrin-containing compound. Other classes of host molecules are crown ethers37–40 and macrocyclic polyphenols. which possess functional properties suitable for several applications such as drug delivery. Adding MgCl2 or CaCl2 . which undergo self-processes. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. hard systems. Using DLS.49–51 Following from the fact that the properties of the block copolymers can be controlled by both the primary structure of the peptide and the size and properties of the synthetic polymer.54. Some are valuable for their curing properties such as antibiotic. DLS.59 investigated a peptide that mimics the repetitive GGX motif of the spidroin silk protein.66 In this regard. (VPGVG)4 and (VPAVG)4 . which is relatively large compared with those obtained with “synthetic” surfactant systems. in water. the oligopeptide adopting an α-helical structure as proven by CD.30–34 Munteanu et al. DLS revealed that the copolymers form spherical structures of 100–200 nm. Pechar et al. Recent progress in this field is further exemplified by the contribution of “click” chemistry. Owing to its unique structure and high mechanical performance. Incubation of peptide-grafted HPMA copolymers.60 presented a detailed analysis of the dynamic properties of entangled semiflexible.43–48 Interest in hybrid materials based on block copolymers of natural polymers (peptides and proteins) and synthetic polymers has increased in recent years. proline (P). elastin-like polypeptides are popular due to their ability to mimic the main extracellular matrix protein that is responsible for the reversible elongation and contraction of various tissues in vertebrates. antifungal. 42 The latter have been successfully used for the design of a variety of receptor molecules showing complexation properties.57 investigated the self-assembly of PEG covalently linked to an oligopeptide. Supramolecular Chemistry: From Molecules to Nanomaterials. while some are secreted by cells into the growth medium. A pair of random coil peptides was also designed to reveal the role of the coiled-coil grafts. which are appealing and versatile structures for the design of inclusion complexes with hydrophobic guests. 55 The composition of those block copolymers is dominated by small hydrophobic amino acids. Yang et al. Shukla et al. such as alanine (A).52. This article is  2012 John Wiley & Sons.41. supported by AFM (atomic force microscopy) and TEM. Both the presence of the terminal hydrophobic Fmoc group and the hydrophilic poly(ethylene glycol) (PEG) chain in the copolymers were shown to play a crucial role in their self-associative behavior. Imura et al. these tailor-made block copolymers can self-assemble into novel supramolecular structures. Similarly. Zhou et al. showed that. Evidence that the metal ions and peptide interaction was obtained when ethylenediaminetetraacetic acid (EDTA) was added. glycine (G). the formation of a mass of nanorod bundles was observed. and valine (V).61 investigated the self-process mechanism of “natural” glycolipid biosurfactants using DLS. thread-like micelles selfprocessed by cationic surfactants such as cetylpyridinium chloride or cetyltrimethylammonium bromide subsequent to the addition of sodium salicylate in aqueous solution. leading to reversible formation of supramolecular thermoresponsive assemblies. These results correlated with the changes in the secondary structure of the peptide grafts as measured by CD spectroscopy. DLS was used to investigate the host–guest interactions and the self-assembly properties of the resulting complexes. Ltd.35. The associative behavior of the peptides and their PEG derivatives was demonstrated by DLS. 36 synthesized cyclodextrin-containing compounds to act as molecular receptors as well as supramolecular building blocks. at higher concentrations. These can be integral components of tissues. a powerful strategy that enables the modular assembly of new molecular entities. 53 In this context. into sponge-like structures with water-channel radii of 50 nm. addition of a competing peptide or a denaturant to the self-assembled hydrogels resulted in partial disassembly or collapse of the structure. such as silica-containing systems. The LS results were consistent with the theoretical model of dynamical coupling between the concentration fluctuations and the dissipative properties of the system.1002/9780470661345. Similar to self-assembling synthetic and natural polymers. The two distinct pentaheptad peptides acted as physical crosslinkers through the formation of antiparallel coiled-coil heterodimers. DOI: 10. or even anticancer agents. Ltd.4 Techniques most important molecular receptors studied in supramolecular chemistry are cyclodextrins.58 studied the kinetics of the self-processes underlying the formation of a hybrid hydrogel system resulting from the organization of hydrophilic synthetic N(2-hydroxypropyl)methacrylamide (HPMA) polymer backbones modified by oppositely charged peptide grafts. spider silk remains an intriguing natural polymer.62–65 It was observed that mannosylerythritol lipids self-assemble into large unilamellar vesicles and. Polymers are the higher molecular weight analogs of surfactants and lipids. Ltd. which still focuses research efforts. which were coupled via their N-termini with activated semitelechelic poly(ethylene glycol) O-(N-Fmoc-2-aminoethyl)O  -(2-carboxyethyl)undeca(ethylene glycol) (Fmoc-PEGCOOH) to yield monodisperse Fmoc-PEG-peptide diblock copolymer. DLS was performed in combination with rheological measurements in order to investigate the dynamic properties of the system. unique and complex molecular structures arecomposed of several chiral centers that are known to be molecularly engineered by microorganisms probably evolved into these sophisticated glycolipid biosurfactants self-assemblies. Online  2012 John Wiley & Sons. Koga et al.56 synthesized short polypeptides. which induced the disruption of the bundles.

69 carefully analyzed the synthesis of the silicalite-1 zeolite. several stimuli influence the properties of supramolecular systems and their reactivity. These observations revealed a complex. cetyltrimethylammonium bromide acted as a lyotropic template. 5 A most sophisticated design of constituent molecules is required to achieve precisely controlled and well-defined supramolecular systems that can respond to external stimuli such as pH or temperature. Using DLS.smc039 . which are held together by noncovalent interactions (mainly London forces).1002/9780470661345. self-assembling copolymers in aqueous media under mild conditions without protection/deprotection chemistry steps. with inherently tunable properties for stimuli responsiveness. The direct synthesis of functional. In combination with DLS. the morphology and mesostructural ordering. whereas an amphiphilic block copolymer stabilized the droplets. or porous flakes were observed and analyzed by DLS. For this purpose. The self-diffusion coefficient provided a way to estimate the equivalent hydrodynamic radius.1 SUPRAMOLECULAR REACTIVITY AND DEVICES Supramolecular reactivity In contrast to the interactions between at least two molecules being referred to as molecular reactivity. in particular. The copolymerization of PVL (poly(vinyl laurate)) and PNIPAM (poly(N-isopropylacrylamide)) resulted in a reduced and broadened lower critical solution temperature (LCST) of the copolymer solutions compared with a solution of PNIPAM alone. Ltd. Ltd. remains a current challenge that focuses research efforts on developing stimuli-responsive delivery systems such as micelles Supramolecular Chemistry: From Molecules to Nanomaterials. and post-LCST aggregation.DLS in supramolecular materials chemistry undergo self-processing. when raising the temperature to 40 ◦ C (above the lowest critical solution temperature of the NIPAM block71 ) while maintaining the pH of the solution at 12. can be tuned. As DLS is particularly suited to the characterization of growth or reduction in size of a reacting assembly. as well as the porosity. Schiller et al. which should be considered when modeling the zeolite assembly process. which is synthesized from tetrapropylammonium hydroxide. polydisperse mixture of particles present at the onset of the zeolite formation. NMR (nuclear magnetic resonance) provides an indirect proof of supramolecular aggregation. whereas DLS yields the size and size distribution of the supramolecular system. address this major goal. either porous particles. Naik and Sokolov68 described the synthesis of micrometer-sized nanoporous and mesoporous silica spheres using disodium trioxosilicate (as an economical silica source) and cetyltrimethylammonium chloride as the structure-directing agent. some examples of supramolecular systems for which this technique was used to analyze the supramolecular reactivity of the ensemble are given in the following. Figure 3 clearly shows the dimensions of the two different supramolecular structures. coil–globule transition at the LCST. and water. They demonstrated that the diblock copolymer can form normal and inverse micelles in aqueous environments by taking advantage of the stimuli-responsive characteristics of both building blocks to pH and temperature. lowering the solution pH to 2 at ambient temperature resulted in the formation of aggregates in which the VBZ block forming the core is now hydrophobic. The mechanism of the sphere formation was studied by DLS and further supported by AFM. Over the course of the synthesis. tetraethylorthosilicate. Aerts et al. supramolecular reactivity refers to interactions of an ensemble of molecules. however. which are schematized in Figure 4. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons.72 synthesized poly(N-isopropylacrylamide-covinyl laurate) copolymers. either hydrophilic or hydrophobic. it was shown that. selfassembling copolymers. Two stages were found: in a first step. By varying the amount of block copolymer. Time-dependent DLS measurements demonstrated that the temperature-induced transition of the copolymers is divided into three stages: pre-LCST aggregation. that is. Online  2012 John Wiley & Sons. However. Conversely. 20–30-nm size primary particles were formed. 3 3.70 described the characteristics of a doubly responsive diblock copolymer composed of N-isopropylacrylamide (NIPAM) and 4-vinylbenzoic acid (VBZ) monomers. causing a pronounced aggregation in solutions. The faster process corresponds to collective particle diffusion. capsules. a few hundred nanometers in size. Cao et al.67 prepared mesoporous silica particulate structures by combining cooperative self-assembly in inverse miniemulsion with a sol–gel process. Ltd. Since the information determining structure formation and interaction of the resulting assembly is encoded in their molecular configuration. which then fused together to form micrometer-sized mesoporous spheres. This article is  2012 John Wiley & Sons. Therefore. supramolecular selfassembly results in nanosized species that are composed of a hydrophobic NIPAM core stabilized by a hydrophilic VBZ corona. DOI: 10. which is commonly used for the fabrication of materials from a solution of chemical precursors of an integrated network (or gel ) of either discrete particles or polymers. 1 H NMR spectrometry was used to assign the hydration of the charged block of the polymer depending on the pH. Depending on the reaction time. Such amphiphilic block copolymers are composed of at least a permanently hydrophilic block and a “smart” block. Lowe et al. which facilitates the formation of hydrophobic microdomains far below the LCST. stabilized by the hydrophilic NIPAM block. The kinetics of the reaction revealed two diffusive processes and polydispersity in size and shape arising from a slower process.

which can be used to trigger the release of an encapsulated hydrophilic guest molecule in a device to transport hydrophilic molecules and release them with such a pH trigger.75 synthesized short amphiphilic calixarene molecules with a small hydrophilic part consisting of amine ethers or amine alcohols. Ltd. Dendrimers have attracted increasing attention in recent years because of their unique structure.76–83 Poly(amidoamine) (PAMAM) dendrimers are monodisperse.8 nm 10 D h = 66. a relatively long part of the macromolecule needs to be influenced in order to change the supramolecular reactivity.84 showed the spontaneous supramolecular complexation of amine-terminated PAMAM dendrimers with an anionic surfactant. Online  2012 John Wiley & Sons. Using DLS. which—in addition to the size and aggregation behavior identified by DLS—yielded the thermodynamic properties of the system. Ltd. This article is  2012 John Wiley & Sons. which was introduced in the last step of the synthesis.1002/9780470661345. such as binding energies between the SDS and the dendrimer. natural blocks such as oligonucleotides. and nanoreactors. VBZ Low pH High T Low T High pH NIPAM Figure 4 media. These results were further confirmed by isothermal titration calorimetry (ITC) measurements.%) 25 20 pH 12 T = 25 °C pH 2 T = 25 °C 15 D h = 15. D h (nm) Figure 3 Hydrodynamic size distribution of self-assembled poly(NIPAM-block -VBZ) block copolymers at pH 12 and T = 25 and 50 ◦ C.smc039 . Since. where PNIPAM is hydrophobic at acidic pH and at high temperatures. Proposed structure formation of poly(NIPAM-block -VBZ) diblock copolymer in normal and inverse micelles in aqueous Supramolecular Chemistry: From Molecules to Nanomaterials. Further increasing the chain length induced the collapse of the vesicles into spherical micelles.7 nm 5 0 1 10 100 1000 Hydrodynamic diameter. Owing to their intrinsic sensitivity to the properties of the surroundings and their ability to undergo specific interactions such as receptor–ligand recognition. when the self-assembly of the PLL-based copolymers is induced in basic media and at low temperatures. sodium dodecyl sulfate (SDS). interesting properties. DOI: 10. They observed a strong electrostatic interaction between the SDS and the amine termination of the PAMAM dendrimers when the pH was below 7. Ltd. they self-assemble into structures in which the PLL is the hydrophobic block. gene therapy. they were able to show a decrease in the size of the supramolecular aggregates with decreased pH and SDS concentration. The binding of SDS induced a physical hydrophobic modification of the outer surface of the PAMAM dendrimer.70 and vesicles. with polymers. and polysaccharides are being investigated as constitutive segments of self-assembling block copolymers. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons.6 Techniques 30 Mean (vol. which promoted the formation of PAMAM/SDS supramolecular complexes.73 polypeptides. they were able to demonstrate the formation of vesicles and observed different self-association behaviors by merely changing a short part of the hydrophilic segment. They showed that PNIPAM-b-PLL copolymers self-assembled into micelle-like aggregates. In contrast. DLS was combined with SEM (scanning electron microscope) to analyze the effect of the length of PLL. as well as their potential applications in medicine. Combining DLS and TEM. Lee et al. pH. The calixarene molecules with a small hydrophilic part assembled into well-defined and tunable vesicles that decrease significantly in diameter with increasing hydrophilic chain length.4. and temperature on the LCST of PNIPAM as well as on the resulting properties of the self-assembly of these PLL-based copolymers. catalysis. highly branched polyelectrolytes with ammonium functional groups at the outer rim (primary amine) and at the branch points (tertiary amine).74 synthesized pH-responsive block copolymers composed of PNIPAM and poly(L-lysine) (PLL). The vesicles were also observed transforming into small globular micelles at lower pH. Zhao et al. Wang et al.

capable of aiming the assembly at tumor cells. Mueller et al. that is. They showed that the p-tert-butyl thiacalix[4]arenes functionalized with hydrazide groups are effective extractants of soft metal cations. Another application of supramolecular devices is in the field of diagnostics. The high number of gadolinium complexes. 86 Rare earth-substituted. Vaccaro et al.and interaggregate electrostatic repulsions on the aggregation mechanism. due to its high number of unpaired electrons. Gd3+ . Cu2+ . in its complex Gd-diethylenetriaminepentaacetate (Gd-DTPA). POMs can offer unique functionality to POMs. whereas on decreasing solution pH or increasing ionic strength. Cd2+ ) block elements was investigated by 7 DLS. allows very effective paramagnetic contrast agents to be obtained compared with micelles formed by first-generation monomers.103–105 the rotation of the complex is slow enough to enhance the relaxation value. In the following section. Online  2012 John Wiley & Sons. By assembling complexes with macromolecules100–102 or supramolecular aggregates. 4 SOFT MATTER AND NANOTECHNOLOGY DLS is not only a powerful technique for investigating supramolecular systems but also a very versatile method Supramolecular Chemistry: From Molecules to Nanomaterials. the effect of electrostatic interactions on intra. These POMs have found applications in various fields including catalysis. spherical. 3. {P4 Y8 W43 } macroions were found to form hollow.94–97 especially in medical applications. The blackberry size increased with decreasing pH (lower charge density) or higher acetone content in the mixed solvent (lower dielectric constant).85. In current medical diagnostics. A constant hydrophobic shell thickness was measured by DLS when the average hydrodynamic radius and size distribution depended on the nature of the encapsulated hydrophobic substrate. the formation of bilayer structures was favored. Ltd. The blackberry size lent itself to accurately control either by changing the acetone content in water–acetone mixed solvents or by changing the solution pH in aqueous solution. is the most utilized.1002/9780470661345. Poly(butadiene)-b-poly(ethylene oxide) self-assembles into unilamellar vesicles in dilute aqueous solution. The system was composed of mixed aggregates formed by an anionic monomer containing the DTPAGlu. especially lanthanidesubstituted. electric. in particular. Contrast results from local differences in spin relaxation time along the longitudinal (T1 ) and transverse (T2 ) planes of the main magnetic field applied to a specimen. such as excellent luminescence properties.99 synthesized novel p-tert-butyl thiacalix[4]arenes functionalized with hydrazide groups at the lower rim of the cone and the partial cone in the 1. without affecting the morphology of the self-assembly. Stoikov et al.2 Supramolecular devices The preparation of nanoscale architectures is critical to the advances in the development of supramolecular devices. each of them characterized by a high relaxation value (r1p = 21. Co3+ . and among these. Ltd. solvent composition influences supramolecular assembly as well as reactivity. which increases the contrast.smc039 . A number of features of the aggregation behavior were revealed by DLS and. Ni2+ . medicine. have been investigated. Pb2+ ) and d-(Fe3+ . Ltd. Polyoxometalates (POMs) belong to a distinct class of oligomeric.3-alternate conformations. or material sciences.DLS in supramolecular materials chemistry In addition to pH. the most frequently used contrast agents are T1 agents. The affinity of the selfassembly to p-(Al3+ . for which the release of active compounds is needed at desired locations. Magnetic resonance imaging (MRI) has become one of the most efficient diagnostic techniques in recent years.0 mM−1 s−1 ).93 studied the self-assembly and formation of “blackberry” type supramolecular structures by a type of yttrium-containing POM (K15 Na6 (H3 O)9 [(PY2 W10 O38 )4 (W3 O14 )]·9H2 O or {P4 Y8 W43 }) macroanions using DLS. Ag+ . single-layer “blackberry” structures in water and water–acetone mixed solvents. Mixed aggregates.98 used DLS to assess the successful embedding of a hydrophobic substrate into the hydrophobic membrane of block-copolymer vesicular structures. formed either by the monomer or by the DTPAGlu derivative associated with the chelating agent or in its complex form.87–92 Mishra et al. All the p-tert-butyl thiacalix[4]arene derivatives with hydrazide fragments were able to form nanoscale aggregates but did not show selfassociation abilities without complex formation with metal cations. a derivative of DTPA and an uncharged monomer containing the bioactive peptide C-terminal cholecystokinin octapeptide (CCK8) amide. The complex stoichiometry depended on the receptor configuration. ultraminiature machine components. examples of such supramolecular devices characterized by DLS are given.104 Micelles and bilayer structures were detected at physiological pH.106 studied in detail a supramolecular system with potential application as a tumor-specific MRI contrast agent using DLS. which are functional at the nanoscale. Imaging agents need to be used in order to improve contrast. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. whereas the blackberry size changed in response to the change of external conditions. structurally well-defined multinuclear complexes with interesting optic. DOI: 10. Release of compounds from delivery systems is currently a major field of research. and magnetic properties relevant to many fields. This article is  2012 John Wiley & Sons.

They determined the radius of gyration and the fractal dimension from the scattered intensity patterns of SLS and X-ray scattering. Working on soft matter systems usually raises important issues that can be answered by the use of DLS.108 This shows that the polydispersity of the primary particles in the silica grades does not influence the comparability of structural and diffusional analysis of pyrogenic silica. contribute to the correlation function and thereby to the particle sizing. which is below the real value. Furthermore. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. by the use of DLS. such as flexible linear or branched polymer samples. The motions of the branching points are coupled with the spring–bead relaxations of a linear chain and can have a noticeable effect on the dynamics of the short chains connecting the two branching units. either repulsive or attractive. which had been degraded in an alcohol suspension by adding different amounts of concentrated HCl. which can help predict the stability and a range of macroscopic properties of colloidal systems. provide a critical challenge for LS experiments. The hydrodynamic aggregate radii were obtained from multiangle DLS. An especially interesting topic is the use of DLS with nonspherical or noncompact objects. and rheological properties. such as SANS and SAXS in characterizing soft matter. Measurements in solutions with gradient salt concentrations can therefore be helpful in finding the optimum salt concentration to screen electrostatic interactions and therefore slow down the diffusion speed and Supramolecular Chemistry: From Molecules to Nanomaterials. The branching units probably influence the relaxation processes. The electrostatic interactions. we cover some of these aspects to demonstrate how broad the field of applicability of DLS is. The degraded starch was studied in an aqueous solution of 0.5 M NaOH as a mimic of randomly hyperbranched polymeric nanoparticles.smc039 . They showed that. one may wonder whether an average diffusion coefficient still has any physical meaning. K¨atzel et al. a diffusion coefficient distribution should be given. such as polymer segment motion. in that the DLS equipment is relatively inexpensive. Because of this effect. in which internal modes of particle mobility. Ltd. Particle size and particle size distribution (PSD) of dispersed systems affect key properties such as surface area. it is possible to directly obtain the Z-average diffusion coefficient of a polydisperse polymer sample. An additional factor exists in solutions of finite concentrations of charged molecules and may increase the diffusion speed—repulsive electrostatic force. the presence of salts in the buffer can screen the intermolecular interactions between the charged molecules. the interconnecting chains between the two branching points were short. which rules out the properties of the investigated starch solutions. as both mean values show good correlation. In these cases. highly branched amylopectin. This example illustrates that DLS is not just a tool for particle sizing by measuring the diffusion coefficient but also yields detailed information about the complex internal dynamics of systems deviating from the simple case of solid spheres. In their publication. Therefore. Such behavior had been predicted by numerical simulations. opacity. or rod-like colloidal particles. Soft matter often deals with charged systems and these systems. This article is  2012 John Wiley & Sons. Quantifiable changes in particle size or PSD provide valuable indications of aggregation or dissociation phenomena. When analyzing polydisperse samples. as the diffusion coefficient distribution for those samples spans an enormous range. These internal modes are not only important for particle sizing but can also provide further interesting information concerning the sample characteristics. Another is the investigation of charged systems and polyelectrolyte solutions such as DNA. Galinsky and Burchard109–111 showed a method of separating contributions stemming from internal modes and particle self-diffusion to the decay rate of the correlation function by using the length scale from the LS experiment. In this section. they can alter the common Zimm–Rouse spectra.112 Their results indicated angular dependence of the apparent diffusion coefficient when the particles possess some internal flexibility.113 The effect was only noticeable if the branching density was high. This chapter is a good example of depolarized DLS measurements used to directly obtain information on rotational diffusion of polydisperse fractal systems. data analysis yields a final result for the particle size.8 Techniques in other fields such as nanotechnology and soft matter. In a more recent study. for polydisperse samples. DLS has advantages over other methods. and does not require special solvents. A normalized plot of the scattered intensity by all nonporous silica grades led to a master curve. Online  2012 John Wiley & Sons. The interpretation of DLS data for pyrogenic silica is of great relevance for technical applications but is hampered by the fractal structure of the aggregates. Ltd. affect chain conformation and influence the diffusion coefficient. 108 investigated a polydisperse fractal system of pyrogenic silica. Mes and coworkers7 compared different methods to analyze the polydispersity of a polymer sample by measuring a series of practically representative SAN copolymers. that is. Working with more complex systems usually present additional factors that must be considered in the correlation function when analyzing the data. does not emit harmful radiation.107. DLS is therefore specially suited to the analysis of high-molecular-mass polymers and is not disturbed by the presence of low-molecular-mass impurities. in particular.1002/9780470661345. They investigated potato starch. reactivity. Ltd. One such issue is the analysis of polydispersity. All samples contained the large. Their DLS results showed a strong angular dependence caused by influences of the rotational diffusion. Characterizing charged particles in solution implies that the particle interactions caused by Coulomb forces must be suppressed. DOI: 10. packing density.

This slow mode is a second-relaxation process found in many DLS experiments performed with polyelectrolyte samples. the data discussed by the authors are not in good agreement with the results obtained with fluorescence spectroscopy. since it may precisely demonstrate the existence of the above-mentioned aggregates. High-molecular-weight calf thymus DNA was dissolved in TE buffer. They presented the data from DLS measurements of DNA macromolecules with chain lengths varying between 700 and 115 000 bp. The other solutions at different salt concentrations (5. Ltd. Ali et al. In this study they investigated the structure and dynamics of DNA solutions in the dilute and semidilute range of concentrations at different temperatures. and the approximate length of DNA was determined by agarose gel electrophoresis to be ∼400 bp. This setup enables an estimation of the size of Brownian scattering centers flowing in microchannels. combining SANS with SLS and DLS. Any residual NaCl in solution was removed by dialysis of the DNA solution against double-distilled water. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. which can precipitate if too much salt is added to the particle solution (salting out). poly(vinylpyrrolidone) (PVP) and PEG. The partial negative charge of the oxygen atom of the amide group present along the polymer backbone and the cationic head group of the drug were found to be responsible for the contraction of the drug–polymer complex. these larger particles served as model dust. They showed that AMT interacted more strongly with PVP than with PEG. This fact can be explained by the attachment of the fluorescent probe to the DNA molecule. Ruf described a different approach to overcome this problem. affecting the diffusivity of the macromolecule. The study of these interactions involving polyions. 117 work in this area. Dust contribution to DLS signals was taken into account by extending the usual scheme by two additional parameters. which made possible a determination of the single particle relaxation process from the polyelectrolyte slow mode. DOI: 10. and it is still not generally understood.smc039 . a surfactant that forms small micelles in aqueous solution. it is important to find an alternative route to remove dust. with certain experiments with rather large samples or with samples that adsorb to the filter membrane. This article is  2012 John Wiley & Sons. thus validating the theoretical estimations experimentally. due to the Poiseuille flow that induces a shear-dependent decorrelation term. Borsali et al. The formation of Supramolecular Chemistry: From Molecules to Nanomaterials. 115 and experimental116. which is mainly interesting if physical removal of the dust is not possible. there is an ultraslow mode. using DLS. with and without the addition of salt. it was possible to deal with the unwanted dust contributions.118 The DLS experiments carried out with DNA at different salt concentrations showed that.122 studied the interaction between an amphiphilic drug. a second population of particles with sizes approximately one order of magnitude larger than the micelles remained in the solution. One major problem in DLS measurements is the contamination of the sample solution by dust particles. Destremaut and coworkers123 established an on-line DLS setup to measure colloidal sizes in a pressure-driven microfluidic flow system. 100. The results obtained from this study indicate a way to use DLS as a tool for the design of a drug delivery system.DLS in supramolecular materials chemistry reveal the actual particle sizes. Online  2012 John Wiley & Sons. These aggregates have been directly observed by other authors119 using cryoelectron microscopy. amitriptyline hydrochloride (AMT). Ltd. However. and neutral polymers. It was shown that. salt-free system by adjusting the salt content. DNA behaves as a worm-like chain for lengths below 10 000 bp. this can be resolved by simple filtration and sometimes followed by a centrifugation step.121 The suggested procedure was tested with octaethylene glycol dodecyl monoether. The Coulomb interparticle interactions were screened by the addition of salt. described the effect of charged interactions on DLS results for the interesting case of DNA in aqueous solution. In the study of Borsali and coworkers. by a relatively simple technique. a special focus on their investigation of hydrophobic polyelectrolytes is of particular relevance. This method treats the dust particle analytically in order to allow particle sizing even for nonfiltered samples. Theoretical arguments were given to underline the difficulties inherent in performing such measurements under flow in a microchannel. suggesting the existence of large aggregates. Ltd. and there has been considerable theoretical114. After filtration and centrifugation. which simplifies the Siegert relation in the case when dust particles are not present. This must be avoided at all costs due to their strong scattering 9 intensity. 250. However. and coions has attracted much interest. This was indicated by a large decrease of the aggregate size with an increase in the AMT concentration. in standard buffer solutions. Thus. in addition to the two well-documented modes (cooperative and slow). In most cases.1002/9780470661345. In another study. counterions. 500. Since the amount of dust present in the system could be adjusted by the total duration of centrifugation. which is thus not comparable with those obtained by LS measurements performed with nonmodified DNA molecules. It can be assumed that these particles are by-products of the synthesis of the surfactants. an offset in the amplitude autocorrelation function and an offset in the intensity autocorrelation function. The normalized intensity and the normalized field autocorrelation function are then related by a quadratic equation.124 They constructed a specific DLS setup around a microfluidic poly(dimethylsiloxane) (PDMS)-based chip. and 1000 mM NaCl) were prepared from the stock. Zimbone and coauthors120 described the combination of DLS and SLS as well as SEM to characterize the conformation of DNA molecules over a wide range of molecular weights. 10. which is a sign of a sequential collapse of the polymer conformation.

providing information on the population and dynamics of the scattering elements associated with cold cataracts. In most cases. DOI: 10. The hydrodynamic size of proteins is thus defined by the molecular weight and arrangement of the polypeptide chains. 126 Proteins can be considered as a complex arrangement of polypeptide chains. the authors demonstrated the robustness of the potential use of DLS as a noninvasive technique for cataract diagnosis. Ltd. applications of DLS to characterize charged protein solutions.130–133 The increase in size and dynamics of aggregate formation can be investigated by DLS by monitoring the kinetics of the size distribution. and the second is the phase separation of the lens cytoplasm into protein-rich and protein-poor phases. polydispersity. However. analytical ultracentrifugation. There are two molecular mechanisms that explain the increase in the intensity of scattered light by cataractous protein lenses.1002/9780470661345. it appears to be an extremely valuable method for the characterization of biological systems in combination with microscopy techniques such as TEM or SEM. This structure is stabilized by a fine balance between hydrogen bonds and electrostatic and hydrophobic interactions providing a large degree of flexibility to the system. The thermal stability of proteins was measured in this way by DLS. The first is aggregation through the formation of high-molecular-weight aggregates. monitoring the hydrodynamic size of a protein is one way of observing its stability under native or denaturing conditions.128 A method of size estimation of the start aggregates has been developed based on the construction of graphs of LS intensity versus hydrodynamic radius. several factors can influence this functional structure. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. Markossian and coauthors127 studied the thermal denaturation and aggregation of rabbit muscle glyceraldehyde3-phosphate dehydrogenase (GAPDH) using differential scanning calorimetry (DSC). Ltd. which increases when both the protein aggregates and the monomers are present in the experimental probe at the same time. and therefore the size of the Supramolecular Chemistry: From Molecules to Nanomaterials. fast dynamics in soft matter systems. and the sample can be recovered for subsequent analyses. accurate.2). since this could significantly influence the results. DLS. and attention must be paid to the refractive index of plastic plates that might differ for different kinds of plastics. The results directly indicate that the initial stage of thermal aggregation of the proteins is the stage of start aggregate formation. just as any other charged molecules in solution. resulting in the absence of a reliable parameter for the cataract index. and pH.134–137 In this case. However. which need powerful near-infrared lasers (830 nm). 633 nm). more generally. or. There are several types of experiments for which DLS is already well established as a technique. phase transition. The authors demonstrated that the process arises from the matching of light wavelengths with the spatial dimensions of fluctuations. this miniaturized device dedicated to DLS measurements is a promising tool to study dynamics on the subsecond timescale. could face certain difficulties in subsequent data treatment. Online  2012 John Wiley & Sons.129 presented a DLS study of the phase separation of the ocular lens emerging on cold cataract development. Hence.smc039 . and SANS. and analytical ultracentrifugation. temperature. these newly developed techniques use He–Ne lasers (red light. This tool might be particularly interesting for investigating nanoparticle synthesis. The intensity of the autocorrelation functions of the lens protein was analyzed with the aid of two methods. which can thus be detected at very low concentration. the results of particle sizing are shifted toward the size of the aggregate.125. The hydrodynamic radius of the start aggregates remained constant in a certain temperature range and was independent of the protein concentration. Petta et al. DLS is also used as a method to characterize the hydrodynamic properties of globular proteins in electrolyte solutions. there is still a lack of systematic methods other than DLS data analysis in cataract diagnostic. or ionic strength might affect the molecular conformation along with a subsequent change in shape and size. in which DLS analysis is combined with microplate reading or chromatography. Ltd. and simple to perform. The optimal use of these modern devices is therefore hampered by the limitations of DLS (Section 1. SAXS. to find optimum protein crystallization conditions. Thus. This article is  2012 John Wiley & Sons.10 Techniques electrostatic coacervates in a solution of oppositely charged nanoparticles and block copolymers in the microfluidic channel were also studied. The DLS data supported a previously proposed mechanism of protein aggregation that involves a first stage of the formation of the start aggregates followed by their subsequent coalescence. and aggregation factors. Modern devices have appeared. This noninvasive technique is fast. For example. On the basis of the meticulous data analysis described in this chapter. Since the scattered intensity is proportional to the molecular radius to the power of six. which organize in a unique threedimensional structure. the scattering intensity. In contrast to conventional biomedical techniques. DLS is used as a prescreening tool to identify critical parameters of the process such as the hydrodynamic size. However. DLS is an established technique for protein characterization. 5 SUPRAMOLECULAR ASPECTS OF CHEMICAL BIOLOGY Even if DLS does not provide the visualization of the particles under investigation. these commercial systems for particle sizing operate at a single detection angle (90◦ ) of the scattered light by the protein solution.

This article is  2012 John Wiley & Sons. polydispersity.DLS in supramolecular materials chemistry particles. is monitored as a function of temperature. was then exemplified through nonexhaustive yet representative examples. TEM. Oxford University Press. the assembly of two proteins of the same size does not yield aggregates with a size that is twice that of the monomers because of the changes of molecular conformation. 2. with this normally happening when the particle size of one population is. the peak of scattering intensities should be resolved. This article was published in the Supramolecular Chemistry: From Molecules to Nanomaterials in 2012 by John Wiley & Sons. we wish to highlight the major limitations of the technique to prevent misuse. the self-assembly of low-molecular-weight analogs of amphiphilic block copolymers. DLS. Huntington’s. Aside from demonstrating the broad field of applicability of DLS. In most cases. Oxford. The changes in size in self-assembled structures from either synthetic or hybrid copolymers in response to changes in temperature. Owing to their biological function and current use in the medical field. was also cited. For example. Ltd. and type II diabetes. 4. Interactions of proteins in solution can be investigated by DLS as well. G. tertiary. An Introduction to Dynamic Light Scattering by Macromolecules. Proc.142 have used DLS to study the self-assembly of a microbial siderophore. we have exemplified the use of DLS in characterizing the properties of supramolecular structures resulting from the assembly of synthetic and hybrid copolymers as well as supramolecular and natural polymers. W. Parkinson’s. 2007. which increases the scattering intensity.141 studied the changes in secondary. in the wide field of supramolecular materials chemistry.1002/9780470661345. Academic Press. Schmitz. These findings are expected to provide a new way of considering amyloid fibrillation and threat related diseases. Soc. 186–203. 219(1137). being the latter often responsible for human diseases. pH. which is the process of the conversion of individual protein molecules into a macroscopic crystal in solution. In this context. It was shown that DLS and SLS could serve as tools for monitoring the thermodynamics and kinetics of protein crystal growth. 3. Wilson143 described DLS as a useful tool to study protein crystallogenesis. However to distinguish between protein assemblies and monomers. Dynamic Light Scattering: The Method and Some Applications. for which the achievement of a comprehensive understanding of the underlying biological mechanism would enable the diagnosis and development of successful treatments.smc039 . for globular proteins. Berlin. Sch¨artl. Taylor. Prior to the description of the use of DLS to monitor the reactivity of supramolecular systems. DOI: 10. such as for target release or extraction. Depending on the concentration of Fe(III). the bulk formation of mature aggregates is subsequent to a midpoint transition of the aggregation of insulin. Sci. S. or existing interactions in the system under investigation as well as careful sample preparation to avoid the presence of dust are essential for accurate particle sizing. Springer. self-assembled from the siderophore. Liu et al. 1990. monitoring the polydispersity index instead of the hydrodynamic radius can be more powerful in the case of such an analysis. DLS is not only a powerful technique for characterizing soft matter but is also relevant for the investigation of hard systems as shown by the characterization of silica-based ensembles. 1953. 6 CONCLUSIONS Our initial aim for this chapter was to review the basic principle of LS with reference to excellent publications on the theoretical background. and terahertz absorption spectroscopy demonstrated a transformation of the native insulin structure from the α-helix into the β-sheet structure prior to aggregation. from micelles to vesicles. Supramolecular Chemistry: From Molecules to Nanomaterials. different morphologies. or solvent composition of the surroundings. DLS appears to be valuable in following the kinetics of chemical reactions and assessing the reactivity of the supramolecular systems. Brown. Ser. surfactants and lipids. However. and quaternary structures of amyloid proteins. 11 Prior knowledge of the size. Phys.138–140 Assembly and fibrillation of amyloid proteins are believed to play a critical role in various human diseases. on average. Ltd. or on complex formation can be assessed by DLS. Online  2012 John Wiley & Sons. which is one in the A–E marinobactin series that facilitates Fe(III) acquisition by the bacteria through coordination of Fe(III) with the marinobactin head group. Heidelberg. Since an increase in the polydispersity index reflects changes in PSD. a twofold increase in the particle size means a sixfold increase of the molecular weight. such as protein crystallization and aggregation. The use of DLS. 1993. A: Math. DLS was also shown to be particularly suited to characterize mechanisms involved in biological systems. Light Scattering from Polymer Solutions and Nanoparticle Dispersions. Ltd. two times larger than the other. With the self-assembly of copolymers being an active field of research. As shown by thioflavin T fluorescence binding. W. The use of DLS further demonstrates the functional supramolecular assembly of devices with a high potential for application as ultraminiaturized machine components that function on the nanoscale. The different sizes and shapes of the systems were easily assessed by DLS and supported by SANS. Heatinduced melting of protein causes molecule denaturation and subsequent aggregation. we referenced conventional self-processed systems at first. K. REFERENCES 1. London. including Alzheimer’s. that is. Owen et al.. R.

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