References

Introduction
Gold nanorods 3D-supercrystals as surface enhanced Raman scattering
spectroscopy substrates for the rapid detection of scrambled prions
Ramn A. Alvarez-Puebla
a
et al. PNAS (2011)
Supercrystal Characterization
Conclusions
Results: Prion Peptide Detection
in Bovine Serum
a
Departamento de Qumica-Fsica and Unidad Asociada
Consejo Superior de Investigaciones Cientificas
Universidade de Vigo, 36310 Vigo, Spain
3D-Supercrystal Fabrication
Results: Prion Peptide Detection
in Human Plasma
The biologically active prion, PrP
C
(A), contains an
alpha helix, which is converted into a beta sheet in
the scrambled, infectious type, PrP
SC
(B).

SERS Spectra:
(C) PrP
C
characterized by three peaks around
1410 cm
-1
as well as a single peak at 762 cm
-1

(D) PrP
SC
A peak is seen at 762 cm
-1
, but
instead of a triplet, a single peak is seen at
1448 cm
-1

(E-F) all contain a 1:99 ratio of PrP
SC
: PrP
C
, but
in every case the single peak is seen, which
indicates all PrP
C
is converted to PrP
SC


Total molar concentration of prion:
(E) 10
-6
M (F) 10
-7
M (G) 10
-8
M
(H) 10
-9
M (I) 10
-10
M (J) Bovine serum
Prions detectable at 10
-10
M total prion
This work Previous work
3
Prion-spiked human blood (b) is not distinguishable from
normal human blood (a)
However,
Prions can be detected in human serum (d) when
compared to normal human serum (c)
Prion presence is especially detectable (e) after
background plasma (c) has been subtracted
(f) is included as a reference for the pure PrP
SC
spectra

SERS Spectra of Human Samples
Scrambled prions can be detected at the
attomol regime in human plasma
High Density of Au NRs is Achieved
by Modified Seeded-Growth Method
Importance
SERS
High enhancement factors are essential for
detecting molecules at extremely low concentrations
Reproducible fabrication of hot spots with
minimum spot-to-spot variability is necessary

Prions A detection method is critically needed
Mad Cow Disease has resulted in the slaughter of
4.4 million cattle in the U.K.
2

Creutzfeldt-Jakob disease in humans is
transmissible, incurable, and fatal, which demands
early detection for its containment and treatment
Supercrystals are Easily Reusable
After plasma cleaning, the SERS spectra for 1NAT
shows no carryover from benzenethiol (BT), indicating
that supercrystals can be reused and thus are
promising in terms of cost effectiveness
Supercrystal SERS Intensities are Greater than
Competing Au NR Varieties
Au NRs form supercrystal islands if
allowed to dry in a moist environment
for 24hrs, aided by their narrow size
distribution ( ~ 3%)
TOP: SERS intensity maps at 1072 cm
-1
for
benzenethiol (BT) at 3 laser excitation wavelengths

BOTTOM: SERS spectra for each excitation
wavelength (left). Gold rod supercrystals (Au-RSC,
this work) are superior to other established Au NR
varieties including ultralong nanowires, single rods,
parallel NRs and fractally aggregated NRs (right).
Surface Enhanced Raman Scattering
- Analytical method for molecule detection based on
visible / near infrared excitation of plasmons on the surface
of a metal substrate, especially at junctions or hot spots
- Plasmon excitation can result in the generation of field
enhancement factors (EFs) on the order of 10
8
, enabling
detection down to single molecules
Prions
- Proteins found in two forms: 1) PrP
C
is naturally
occurring and biologically functioning, 2) PrP
SC
is
misfolded and infectious; its folding promotes the transition
of PrP
c
to PrP
SC
in a cascade that leads to plaque
formation in neural tissues and subsequent death
SERS: inelastic light scattering at a hot spot
1
(left).
Prions are responsible for Mad Cow Disease
2
(right)
Using a modified seed growth method, Au NRs were
formed with a narrow size distribution
Drying these Au NRs slowly created Au supercrystals that
had superior enhancement factors compared to other Au
NR varieties
Au supercrystals were shown to be extremely robust to
cleaning and therefore easily reusable
Prion peptides were detected in bovine serum and human
plasma at concentrations as low as 10
-10
M
This work demonstrates that label-free prion detection
in relevant biological media is achievable, paving the
way for more effective disease diagnostics
1. Ding, S.-Y. et al. SERS: General Introduction. Encyclopedia of
Analytical Chemistry (2006)
2. Brown, D. The recipe for disaster that killed 80 and left a 5bn bill.
The Telegraph (2000)
3. Nikoobakht, B. & El-Sayed, M. A. Preparation and Growth Mechanism
of Gold Nanorods (NRs) Using Seed-Mediated Growth Method. Chem.
Mater. 15, 19571962 (2003).
Modified protocol: The growth solution consisted of mM
concentrations of CTAB, Au (III) ions, AgNO
3
, and ascorbic acid.
A large excess of seed solution (12mL) was added to the growth
solution to grow NRs for 2 hrs, after which additional ascorbic
acid was added slowly to continue growth (step 1->2 below). NRs
then crystallize into colloidal 3D crystals if allowed to dry for 24hr
in a moist saturated, moist atmosphere (step 3).