Modifications in the nanoparticle-protein

interactions for tuning the protein

adsorption and controlling the stability of
complexes
Cite as: Appl. Phys. Lett. 118, 153701 (2021); https://doi.org/10.1063/5.0046745
Submitted: 06 February 2021 . Accepted: 21 March 2021 . Published Online: 13 April 2021

Sugam Kumar, Debasish Saha, Shin-ichi Takata, Vinod K. Aswal, and Hideki Seto

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Appl. Phys. Lett. 118, 153701 (2021); https://doi.org/10.1063/5.0046745 118, 153701

© 2021 Author(s).
 Applied Physics Letters ARTICLE scitation.org/journal/apl

Modifications in the nanoparticle-protein

interactions for tuning the protein adsorption
and controlling the stability of complexes
Cite as: Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745
Submitted: 6 February 2021 . Accepted: 21 March 2021 .
Published Online: 13 April 2021

Sugam Kumar,1 Debasish Saha,1 Shin-ichi Takata,2 Vinod K. Aswal,1,3,a) and Hideki Seto4

AFFILIATIONS
1
Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai 400 085, India
2
J-PARC Center, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195, Japan
3
Homi Bhabha National Institute, Mumbai 400 094, India
4
J-PARC Center, High Energy Accelerator Research Organization, Tokai, Ibaraki 319-1106, Japan

a)
Author to whom correspondence should be addressed: vkaswal@barc.gov.in

ABSTRACT
We report the pathways to suppress or enhance the protein adsorption on nanoparticles and thereby control the stability of the
nanoparticle-protein complexes with the help of selective additives. This has been achieved by tuning the electrostatic interaction between
the nanoparticles and proteins, in the presence of surfactant and multivalent counterions. The preferential binding of the proteins with the
surfactant and multivalent ions induced charge reversibility of nanoparticles can lead to adsorption of an otherwise non-adsorbing protein
and vice versa. The findings are demonstrated for anionic silica nanoparticles and two globular proteins [lysozyme (cationic) and bovine
serum albumin (BSA) (anionic)] as model systems, in the presence of two ionic surfactants [anionic sodium dodecyl sulfate (SDS) and cat-
ionic dodecyltrimethylammonium bromide (DTAB)], and ZrCl4 as multivalent salt. Small-angle neutron scattering with the unique advan-
tage of contrast variation has been used to probe the role of individual components in the multi-component system. It is shown that the
non-adsorbing behavior of BSA with silica nanoparticles changes into adsorbing in the presence of oppositely charged DTAB surfactant,
whereas the strong adsorbing behavior of lysozyme on nanoparticles modifies to be non-adsorbing in the presence of oppositely charged SDS
surfactant. The presence of multivalent counterions (ZrCl4) leads the charge reversal of the nanoparticles, transforming the lysozyme from
adsorbing to non-adsorbing, and no significant change in the behavior of BSA. The results presented can find potential applications in the
field of nanobiotechnology.
Published under license by AIP Publishing. https://doi.org/10.1063/5.0046745

The integration of nanoparticles with proteins has a prime inter-
est in the field of nanobiotechnology, where these complexes are aimed
to be utilized in different applications such as targeted drug delivery
and biosensing.1–5 The proteins have the tendency to immediately
cover the nanoparticles in the biological milieu to form the protein
corona (a tightly packed layer of protein molecules) around the nano-
particles. However, the uncontrolled formation of a protein corona
around nanoparticles can limit applications due to prompt clearance
from the systemic circulation by macrophages, invoking the need of
controlled and selective protein adsorption.1,6,7
The protein adsorption on nanoparticles is governed by several
interactions, such as hydrogen bonding, electrostatic complexion,
covalent bonding, and hydrophobic attraction, depending on the phys-
ical nature of the nanoparticles and protein.8–10 The functionality of
the conjugate and its response toward the biological target is governed
by the adsorbed layer of the proteins.1,10,11 The adsorption of human
serum protein on iron oxide or calcium carbonate nanoparticles is
shown to be improving the uptake of the complex by cancerous cells
in comparison to bare nanoparticles.12,13 It has also been shown that
the interaction of gold nanoparticles with different cells is decided by
the type of the protein corona.11,14
However, despite the tendency of the protein to adsorb on nano-
particles, in some cases the repulsive interactions between nanopar-
ticles and proteins (e.g., electrostatic repulsion between similarly
charged nanoparticles and proteins) are so strong that they do not
allow the protein adsorption.15 For example, the adsorption of the
serum protein on the silver nanoparticles is suppressed by introducing
additional steric repulsion between nanoparticles and serum proteins

Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745 118, 153701-1
Published under license by AIP Publishing
 Applied Physics Letters
for reducing the cellular uptake of the nanoparticles and its cytotoxic-
ity.16 The tunability between adsorption and non-adsorption with
respect to some physicochemical parameters can enable the unique
advantages that can be exploited for various purposes.1,10,11,17,18 For
example, the idea of utilizing these complexes as carriers for targeted
drug delivery requires both the adsorption as well as release of the pro-
tein (biomolecules, in general) at different instances.17,18 Similarly, in
an interesting study, adsorption of fusion protein on silica nanopar-
ticles has been utilized as a shield to suppress the adsorption of serum
proteins. The MD simulations carried out in this study suggest that
the electrostatic interaction between the shield protein coated nano-
particles and serum proteins is an important factor in minimizing the
serum proteins’ adsorption.7
The interaction between the nanoparticles and protein also dic-
tates the stability of the system, which is another important parameter
for the application of the nanoparticle-protein complexes.15,19 In the
case of oppositely charged nanoparticles and protein, the strong elec-
trostatic attraction leads to the protein-mediated aggregation of the
nanoparticles.15 The non-adsorption of the protein on the nanopar-
ticles may also give rise to nanoparticle aggregation via protein-
induced depletion attraction between nanoparticles.20 The aggregation
in the nanoparticle-protein systems may be useful in some instances
such as to generate the multi-scale equilibrium clusters,21 however,
most of the time it is undesired as it hinders the interaction of the
complex with the biological environment. Nevertheless, the interac-
tions between nanoparticle and protein may be tuned to gain control
over the protein adsorption on the nanoparticles as well as colloidal
stability of the resulting complexes.1,10,22,23
In this Letter, we have tuned the interaction between nanopar-
ticles and protein to control the adsorption/desorption of proteins on
nanoparticles and colloidal stability of the system using oppositely
charged ionic surfactants and multivalent counterions. Anionic silica
(SiO2) nanoparticles [Ludox HS40 (diameter 16 nm and zeta poten-
tial 60 mV), 40 wt. % dispersion in water] and two globular pro-
teins [anionic bovine serum albumin (BSA) and cationic lysozyme],
which are oppositely charged at pH 7.0 were used as model systems
(Fig. S1 in the supplementary material). The silica nanoparticles are
one of the most widely studied model nanoparticles in the field of
nanobiotechnology due to their several relevant properties such as low
toxicity, high stability, good compatibilities with other materials, and
ability to be functionalized with a range of macromolecules.24,25 The
lysozyme protein adsorbs on the silica nanoparticles due to strong
electrostatic attraction, resulting in protein-mediated nanoparticle
aggregation. On the other hand, BSA protein experiences electrostatic
repulsion with nanoparticles and hence remains non-adsorbing. The
non-adsorbing nature of the BSA gives rise to depletion attraction
between the nanoparticles, which again causes particle aggregation but
at significantly higher concentration, compared to lysozyme.15 Two
surfactants (Fig. S2 in the supplementary material), namely, anionic
sodium dodecyl sulfate [SDS: CH3(CH2)11SO4Na] and cationic dode-
cyltrimethylammonium bromide [DTAB: CH3(CH2)11N(CH3)4Br]
were used as both the surfactants have strong interactions with pro-
teins, and the results are supported by dynamic light scattering (DLS)
measurements. Small-angle neutron scattering (SANS) has been
employed to investigate the nanoparticle-protein systems with suitable
contrast conditions. The unique advantage of contrast variation ena-
bles to probe the role of individual component in such multi-
Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745
Published under license by AIP Publishing
ARTICLE scitation.org/journal/apl
component systems (like in the present study) by suppressing the scat-
tering contribution of others.26,27
The ability of contrast variation in SANS originates due to the
presence of the contrast term (qp – qs)2 in the equation for the intensity
of the scattered neutrons, which can be expressed in terms of differen-
tial scattering cross section as27,28 dX
dR
ðQÞ ¼ UVðqp  qs Þ2 PðQÞSðQÞ
þB, where Q [¼ (4p sinh)/k, 2h is the scattering angle, and k is the
mean wavelength of neutrons] is the scattering vector, U is the volume
fraction of scatterers, and V is the volume of the individual scatterer. qp
and qs are scattering length densities of scatterer and solvent, respec-
tively. If qs is equal to the qp for any component (e.g., nanoparticle) in
the multi-component system (e.g., nanoparticle-protein system), the
scattering from that component will have no contribution, and hence
the SANS data will only be governed by the scattering from the other
component (e.g., protein).26,27 In the present study, the scattering
length density of the solvent is varied by mixing the suitable amount of
H2O (qH2O ¼ 0.56  1010/cm2) and D2O (qD2O ¼ 6.38  1010/cm2)
to match them with the scattering length density of silica nanoparticles
(qsilica ¼ 3.80  1010/cm2).26 P(Q) is intraparticle structure factor and
S(Q) is interparticle structure factor. B is a constant term representing
incoherent background. P(Q) depends on shape and size of the scat-
terer, whereas S(Q) provides the information on the interparticle inter-
actions. It may be mentioned here that the use of D2O instead of the
H2O is known to give rise some minor effects on protein-solvent inter-
actions. However, in the present study, no significant effect of the H2O
or D2O or H2O/D2O mixed solvent was observed on the nanoparticle-
protein interactions, as confirmed by DLS.29,30
SANS measurements were performed at the small and wide-
angle neutron scattering instrument (TAIKAN) at J-PARC, Japan.31
The details of the experiments and data analysis can be found in the
supplementary material. Figure 1 shows the SANS data of 1 wt. % sil-
ica nanoparticles (polydisperse sphere, mean radius ¼ 8.0 nm, supple-
mentary Fig. S3 and supplementary Table S1)26 with 1 wt. % lysozyme
protein (ellipsoidal shape, 2.4  1.4  1.4 nm3, supplementary Fig. S3
and supplementary Table S1)32 in the absence [Fig. 1(a)] and presence
[Figs. 1(b) and 1(c)] of SDS surfactant, in the two contrast conditions,
where (i) all the components are visible (solvent is 100% D2O)
[Fig. 1(b)] and (ii) nanoparticles are contrast-matched (solvent is 62%
D2O in H2O/D2O mixture) [Fig. 1(c)]. The interaction of the anionic
silica nanoparticles with the cationic lysozyme protein (in the absence
of the SDS) leads to the protein-mediated aggregation of the nanopar-
ticles. This is evident as the sample becomes turbid (supplementary
Fig. S4), and the SANS data [Fig. 1(a)] show a linear rise in the
scattering intensity in the low Q region due to the formation of the
fractal-like aggregates.32,33 The data are analyzed by employing P(Q)
of core-shell for the building block (nanoparticle core with adsorbed
protein shell) and S(Q) for the mass fractal-like aggregation (see the
supplementary material). On the other hand, SDS surfactant does not
have any physical interaction with the nanoparticles due to similar
charge of the two components (supplementary Fig. S5).26 The addition
of 5 mM SDS, in 1 wt. % HS40 þ 1 wt. % lysozyme system, leads to the
increase in scattering intensity compared to that in the absence of
SDS, without changing the scattering features [Fig. 1(b)]. The physical
state of the sample also remains turbid (supplementary Fig. S4). The
data hence in this case (1 wt. % HS40 þ 1 wt. % lysozyme þ 5 mM
SDS) are analyzed using the same model, which provides much larger
shell thickness than that obtained for the 1 wt. % HS40 þ 1 wt. %
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 Applied Physics Letters
FIG. 1. (a) SANS data of 1 wt. % HS40 þ 1 wt. % lysozyme along with pure components, (b) SANS data of 1 wt. % HS40 þ 1 wt. % lysozyme with varying concentration of
SDS in 100% D2O (all components are visible), (c) in 62% D2O in D2O/H2O (nanoparticles are contrast-matched), and (d) SANS data of 1 wt. % lysozyme with varying SDS
concentrations.
lysozyme system (supplementary Table S2). This may happen if the
nanoparticle aggregation is driven by the lysozyme-SDS complex
instead of the lysozyme alone. The lysozyme protein also interacts
strongly with the SDS, since lysozyme and SDS are oppositely charged.
The lysozyme-SDS complex forms hexagonally closed packed (HCP)
fiber structures at the mentioned SDS to lysozyme molar ratio
[Fig. 1(d) and supplementary Table S2].34 The lysozyme protein and
SDS are known to form different structures at the different molar ratio
of the lysozyme and SDS.35 When SDS is added in the
HS40 þ lysozyme system, the lysozyme preferably binds with the SDS,
which may be due to the combined dominating effect of the hydro-
phobic and electrostatic interactions in the protein-surfactant system
in comparison to electrostatic attraction alone in the nanoparticle-
protein system.36 The complex is then attached to the nanoparticle
surface and leads to the complex-mediated nanoparticle aggregation.
It should be noted that about 6 mM SDS is required to neutralize all
the molecules of the lysozyme (charge þ 8e.u.) in 1 wt. % solution,
which implies that at lower SDS concentration (5 mM) the lysozyme-
SDS complex still carry positive charge, and therefore is attracted
toward the anionic silica nanoparticles.37 On the contrary, the physical
state of the solution becomes clear, with no signature of the nanoparti-
cle aggregation, on raising the SDS concentration to 15 mM. In this
case, the scattering intensity in the low Q region decreases along with
Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745
Published under license by AIP Publishing
ARTICLE scitation.org/journal/apl
the disappearance of the linearity in the scattering profile. In fact, the
data have typical features of the two-form factors governed scattering
contributions from two different sizes [Fig. 1(b)]. To understand the
system behavior, we have contrast-matched silica nanoparticles with
the solvent [Fig. 1(c)]. In the absence of the SDS, the scattering inten-
sity is quite weak of aggregated core-shell structure, as the contrast-
match points of the silica nanoparticles and the lysozyme protein are
nearly same.38 On addition of 5 mM SDS, the data show strong linear
scattering with a small oscillation at Q 0.035 Å1, which can be
understood from a fractal-like structure formed by the shell-like build-
ing blocks (adsorbed protein shell with nanoparticle core contrast
matched). The shell thickness in this case is consistent with that
obtained in the no contrast-match condition [Fig. 1(b)]. However, at
15 mM SDS concentration the scattering profile shows typical features
of the pure SDS micelles, suggesting the coexistence of SDS micelles
with nanoparticles. To understand whether lysozyme interacts with
SDS or nanoparticles or both, SANS data of 1% lysozyme with 15 mM
SDS are examined [Fig. 1(d)]. The data of 1 wt. % lysozyme þ 15 mM
SDS show typical features of the SDS micelles, suggesting the forma-
tion of transparent and soluble complexes of SDS micelles with
unfolded lysozyme at this concentration.35 These complexes carry an
overall negative charge and comprises a decorated core-shell structure,
where the core is made-up of dodecyl chains while the shell is formed
118, 153701-3
 Applied Physics Letters
FIG. 2. (a) SANS data of 1 wt. % HS40 þ 1 wt. % BSA system with varying concentration of the DTAB in 100% D2O (all components are visible), (b) in 62% D2O in D2O/H2O
(nanoparticles are contrast-matched), and (c) SANS data of 1 wt. % BSA with varying concentration of DTAB.
by SDS head groups along with protein in molten globule state.35 The
charge-charge repulsion between the nanoparticles and SDS-lysozyme
complex (15 mM) does not allow adsorption of the complex on nano-
particles and the complex coexists with individual nanoparticles with-
out any significant interaction. In accordance with this, the data of the
1 wt. % HS40 þ 1 wt. % Lysozyme þ 15 mM SDS system could be ana-
lyzed by combining the scattering contributions from the nanopar-
ticles and SDS micelles (supplementary Fig. S6). The fitted parameters
are shown in supplementary Tables S1 and S2.
So far, our results show that the addition of the SDS in higher
concentration could suppress the strong adsorption of the lysozyme
on the nanoparticles and hence the protein-mediated nanoparticle
aggregation. At this SDS concentration, the protein preferably interacts
with the surfactant via hydrophobic interaction instead of charge
interaction with the nanoparticles, as overall charge of lysozyme is
expected to be neutralized by anionic SDS (section S.3 in the supple-
mentary material).37 Now, we illustrate that the suitable surfactant
(cationic DTAB) can also be used to adsorb (or to enhance the protein
adsorption) a protein (BSA) on nanoparticles (anionic silica), which is
otherwise non-adsorbing, as both protein and nanoparticles are simi-
larly charged at pH 7.
SANS data of 1 wt. % HS40 þ 1 wt. % BSA without and with
DTAB surfactant, in the two contrast conditions, where all compo-
nents are visible [Fig. 2(a)] and nanoparticles are contrast-matched to
the solvent [Fig. 2(b)] are shown in Fig. 2. The scattering data of the
1 wt. % HS40 þ 1 wt. % BSA system show a slight increase in the low
Q region compared to that of pristine 1 wt. % HS40. It is known that
the BSA gives rise to depletion attraction between the nanoparticles at
higher concentrations due to its (the BSA’s) non-adsorbing nature
(arising from the strong electrostatic repulsion between nanoparticle
and protein), high number density and smaller size (SizeHS40: SizeBSA
2.8) compared to nanoparticles.15 Therefore, the low Q scattering
build-up can be attributed to the changes in the interparticle potentials
hence is modeled by a single Yukawa potential taking account of
depletion attraction (supplementary Table S3). On addition of the
DTAB, the scattering intensity increases in the low Q region in both
cases and the samples become turbid (supplementary Fig. S4), sugges-
ting the nanoparticle aggregation. Unlike SDS, the cationic DTAB
interacts with both anionic nanoparticles as well as anionic BSA pro-
tein (supplementary Figs. S1 and S2). Hence, to understand the forma-
tion of the complexes in this system, the SANS data of the DTAB with
Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745
Published under license by AIP Publishing
ARTICLE scitation.org/journal/apl
1 wt. % nanoparticles (supplementary Fig. S7) and with 1 wt. % BSA
[Fig. 2(c)], separately are measured. The physical appearance of the
nanoparticles-DTAB system was turbid while that of the DTAB-BSA
system was transparent (supplementary Fig. S4). It is found that the
DTAB micelles strongly adsorb on the nanoparticles and aggregate
them, due to the micelles are oppositely charged to the nanoparticles.26
On the other hand, DTAB micelles unfold the protein and form a
bead-necklace kind of complex, where the surfactant micelles are
arranged along the unfolded protein chain.39 It is therefore clear that
in the three-component (1 wt. % HS40 þ 1 wt. % BSA þ 5 or 15 mM
DTAB) system, the nanoparticles either interact through DTAB
micelles alone or DTAB-BSA complex.
In order to confirm the actual condition, SANS measurements
were performed at a condition where nanoparticles were contrast-
matched. The SANS data again show the signature of fractal-like
structure with a core-shell kind of building block, where the core
(nanoparticles) is contrast-matched to the solvent.26 The fitted shell
thickness suggests that the aggregation is mediated by the BSA-DTAB
complex, similar to the case of lysozyme and SDS. Therefore, the addi-
tion of the suitable surfactant leads to the adsorption of the protein
which is otherwise non-adsorbing.
In the above cases of nanoparticle protein interactions in the
presence of surfactants, we could achieve the tuning of the protein
adsorption from strongly adsorbing to non-adsorbing for lysozyme
and from non-adsorbing to adsorbing for BSA, via complex formation
with the surfactant, but the proteins lose their native structure. To
avoid this, the multivalent counterions, which are known to anoma-
lously alter the double layer around the nanoparticles as well as pro-
teins, to tune the protein adsorption on the nanoparticles, can also be
employed.40,41 Recently, a multivalent counterions driven re-entrant
phase behavior has been observed in the both nanoparticles and pro-
teins (individually), where these systems undergo a transformation
from one-phase (individual stabilized) to two-phase (aggregation or
liquid-liquid phase separation) and back to one-phase (individual)
with monotonic increase in the multivalent ions concentration.42–46
Such re-entrant phase behavior arises due to excessive condensation of
the multivalent counterions on the nanoparticles/proteins, which in
turn reverse the charge of the nanoparticles/proteins.47 Such excessive
condensation is believed to be arising from the strong ion-ion correla-
tions, prevailing in the multivalent counterions and is usually not
observed for the monovalent ions.48 The emergence of the re-entrant
118, 153701-4
 Applied Physics Letters ARTICLE scitation.org/journal/apl

behavior is beyond the standard theories of the colloidal stability, e.g.,
DerjaguinLandauVerweyOverbeek (DLVO) or Debye H€ uckel
(DH) theories (section S.4 in the supplementary material) and is find-
ing profound applications in physical as well as natural sciences.48–51
We have utilized this ability of the multivalent ions to reverse the
charge of nanoparticles/protein facilitating the tuning of the electro-
static interaction between nanoparticles and the two proteins.
Figure 3(a) shows the charge reversal in the nanoparticles in the
presence of ZrCl4, where the measured zeta potentials for 1 wt. %
HS40 silica nanoparticle solution with varying ZrCl4 concentration are
plotted. At low ZrCl4 concentrations (CZr < 1 mM), the nanoparticles
are stable and having a negative zeta-potential. On increasing salt con-
centration (1 mM < CZr < 2 mM), the nanoparticles aggregate and
zeta-potential could not be measured in this range. Further addition of
the ZrCl4 (CZr > 2 mM) leads to a re-entrant stable phase with a posi-
tive zeta-potential, suggesting the charge reversal in the system.42 The
SANS data of the 1 wt. % HS40 nanoparticles in the presence of
20 mM ZrCl4 and 200 mM NaCl (same ionic strengths) are shown in
Fig. 3(b). As can be observed, the data in the presence and absence of
the salt remain more or less same, and without any signature of the
aggregation, suggesting that the nanoparticles are all stable (supple-
mentary Fig. S8).42
The nanoparticle protein interaction has been tuned by adding
20 mM concentration of the ZrCl4, where the negative charge of both
nanoparticles and BSA changes to positive (as both of these reverse
their charges, supplementary Fig. S9), while that of lysozyme remains
unaltered (in this case, Cl- are the counterions, which unlike multiva-
lent counterions do not adsorb on the lyzozyme due to lower charge
density). As a result, the interaction between nanoparticle and
lysozyme is changed from attractive to repulsive. The SANS data
[Fig. 3(c)] of the 1 wt. % HS40 þ 1 wt. % lysozyme system in the pres-
ence of 20 mM ZrCl4, exhibit no signature of the aggregation, as
observed in the absence of ZrCl4 [Figs. 1(a) and 3(c)]. In fact, the data
almost match with that of the pure nanoparticles in the low Q region,
suggesting the non-adsorption of the protein on the nanoparticles
(also verified by DLS, supplementary Figs. S8 and S10). A small scat-
tering build-up in the high Q region arises from the free protein, which
again demonstrating that the protein mostly remains free in the solu-
tion.32 It should be noted that the presence of the 200 mM NaCl does
not show any significant changes in the scattering data, implying that
the monovalent ions are not able to alter the nanoparticle-protein
interactions.44,47–50 On the other hand, SANS data of the 1 wt. %
HS40 þ 1 wt. % BSA system show similar features as observed without
addition of 20 mM ZrCl4. An increase in the low Q scattering can be

FIG. 3. (a) Zeta-potential of 1 wt. % HS40 nanoparticles as a function of the ZrCl4 salt concentration and the physical appearance of the nanoparticle dispersion with increasing
salt concentration. Insets show the physical appearance of the nanoparticle dispersion with increasing salt concentration, (b) SANS data of 1 wt. % HS40 nanoparticles in
the absence and presence of salts (200 mM NaCl and 20 mM ZrCl4), (c) SANS data of the 1 wt. % HS40 þ 1 wt. % lysozyme without and with salts, and (d) SANS data of the
1 wt. % HS40 þ 1 wt. % BSA system without and with salts. SANS data, presented in this figure, are collected in contrast condition where all the components are visible
(solvent is 100% D2O).

Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745 118, 153701-5
Published under license by AIP Publishing
 Applied Physics Letters
FIG. 4. Schematic showing the nanoparticle-protein interactions in the presence of surfactants (SDS/DTAB) and multivalent ions (Zrþ4) and resultant structures formed at each
step.
observed, suggesting the evolution of the attractive depletion interac-
tion. In this case, the protein adsorption on nanoparticles does not
enhance as both are expected to be charge reversed. This shows that
the suitable multivalent counterions in appropriate concentrations can
be employed to tune the protein adsorption.41
To conclude, the present study shows pathways to tune the
interaction between nanoparticles and proteins and thereby protein
adsorption on nanoparticles as well as colloidal stability of the com-
plexes formed. Figure 4 shows schematics of nanoparticle-protein
complexes formed in the presence of surfactants (SDS/DTAB) and
multivalent ions (Zrþ4). It has been demonstrated that the preferen-
tial binding of the protein with the surfactant and the multivalent
ion driven charge inversion of the nanoparticles/proteins can be uti-
lized to create switching between the protein adsorption and non-
adsorption. Moreover, these parameters can also enable the control
over the undesired protein adsorption and nanoparticle aggregation
in the nanoparticle-protein systems. Since the methodology utilizes
tuning of the electrostatic interaction between the silica nanopar-
ticles and proteins, we believe that the findings are transferable to
the other charged nanoparticles (e.g., metal nanoparticles). In this
regard, other additives such as ionic biomolecules (DNA, lipids),
ionic liquids, and charged polymers, which are able to tune the elec-
trostatic interaction between the nanoparticles and proteins, can
also be utilized. However, different strategies are to be designed for
the systems, where the protein adsorption is facilitated by other
interactions such as covalent or hydrogen bonding. Overall, our
studies can find excellent potential applications in the field of
Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745
Published under license by AIP Publishing
ARTICLE scitation.org/journal/apl
nanobiotechnology for creating tunable nano-biohybrids with
directed response, for predicting and controlling the phase stability
and for producing multiscale equilibrium clusters.
See the supplementary material for details of experiments, data
analysis (procedure and fitted parameters), and additional data.
The neutron experiments at the Materials and Life Science
Experimental Facility at J-PARC were approved by the Neutron
Science Program Review Committee (Proposal No. 2019B0033).
This work was supported by a Grant-in-Aid for Scientific Research
on Innovative Areas (No. JP19H05717: Aquatic Functional
Materials).
The authors declare no conflict of interest.
DATA AVAILABILITY
The data that support the findings of this study are available
from the corresponding author upon reasonable request.
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Appl. Phys. Lett. 118, 153701 (2021); doi: 10.1063/5.0046745 118, 153701-7
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