COMPANY PRESENTATION PRODUCT GROUP NEXTERION

SCHOTT NEXTERION®

NEXTERION Slide P - The slide of choice for antibody microarrays

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

NEXTERION® Slide P

Coating Structure and Probe Attachment Principle

Proprietary polymer, ensuring very low non-specific binding

Glass support

C O

O

N

O

O

Attachedto polymer

NHS-ester

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

NEXTERION® Slide P – Spot morphology

4µg/ml RaG in PPB

20µg/ml RaG in PPB

Marker spots

100µg/ml RaG in PPB

500µg/ml RaG in PPB

1000µg/ml RaG in PPB

1500µg/ml RaG in PPB

1800µg/ml RaG in PPB

Protein Print Buffer only

500µg/ml RaG in PP

100µg/ml RaG in PP

Marker spots

Slide P(30 µg/ml labeled Goat IgG)

Excellent spot morphology on NEXTERION Spot PB

Data obtained using a contact printer

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

NEXTERION® Slide P – Binding capacity

High binding capacity on Slide P – up to 1mg/ml probe concentration

signal intensity at various probe and target concentrations

0

0,1

0,2

0,3

0,4

0,5

0,6

0,7

0,8

4 20 100 500 1000 1500 1800

c in µg/ml IgG RaAG

sig

nal

in

ten

sity

30

6

1,25

0,25

0,05

0,01

Hyb concentration in µg/ml IgG from Goat serum

(probe)

(target)

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Hintergründe aller Oberflächen und Proben

0

200

400

600

800

1000

1200

1400

1600

1800

2000

H B E AL

Inte

ns

ity

Rabbit anti Goat IgG 130PMT

Plasminogen 120PMT

GAPDH 120PMT

Galaktosidase 200PMT

H P E AL

NEXTERION® Slide P – Background

Extremely low backgrounds on Slide P shown with various targets

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Signal to Background aller Oberflächen 30µg/ml 130PMT

0

2000

4000

6000

8000

10000

12000

RaG 4µg/ml RaG 20µg/ml RaG 100µg/ml RaG 500µg/ml RaG 1000µg/ml RaG 1500µg/ml RaG 1800µg/ml RaG 500µg/ml PP8,5

Spottingprobe

Sig

nal

/ B

ackg

rou

nd

Signal/BG H 30

Signal/BG P 30

Signal/BG E 30

Signal/BG AL 30

NEXTERION® Slide P – Signal to Bkg ratios

Superior S/B ratios for slide P

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

NEXTERION® Slide P – Signal to Bkg ratios

Superior S/B ratios for slide P

Vergleich Bindungskapazität; Signal to Background; Osteopontin; 532nm 130PMT

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

1000031

25 625

125 25 5 1

3125 625

125 25 5 1

3125 625

125 25 5 1

3125 625

125 25 5 1

3125 625

125 25 5 1

3125 625

125 25 5 1

Slide H Slide P GenTel Whatman Nexterion NC-W Nexterion NC-D

S/B

rat

io

Anti-SPP1 0,03125mg/ml

Anti-SPP1 0,0625mg/ml

Anti-SPP1 0,125mg/ml

Anti-SPP1 0,25mg/ml

Anti-SPP1 0,5mg/ml

Anti-SPP1 1,0mg/ml

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Spot – Diameter with Next spot PB

• Spot morphology was good on all substrates• But only Nexterion NC-W/D and the FAST slide gave acceptable

spot sizes for higher concentrations of detergents in the print buffer

0

20

40

60

80

100

120

140

Slide H Slide P GenTel Nexterion NC-D Nexterion NC-W Whatman

Sp

otd

urc

hm

es

se

r in

µm

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Summary of Benchmark Testing  NEXTERION®

Slide HNEXTERION®

Slide PFast slide

NEXTERION® Slide NC-W (N)

NEXTERION® Slide NC-D

Path slide

Bind. capacity and signal int. (0,25)

+++ +++ +++ ++++ +++ +

Signal/Background ratio (0,25)

++++ ++++ + ++ +++ +++

Probe target linearity (0,2)

+++ +++ ++++ ++++ ++++ +

Sensitivity (0,15) ++++ ++++ + ++ +++ ++++

Print buffer tolerance (0,15)

++ ++ ++++ ++++ ++++ ++

Storage of printed slides

-20°C -20°C RT RT RT ?

 

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Referenced KTH Paper

Rebecca Rimini, Jochen M. Schwenk, Mårten Sundberg, Ronald Sjöberg, Daniel Klevebring, Marcus Gry, Mathias Uhlén and Peter Nilsson

Validation of serum protein profiles by a dual antibody array approach

Journal of Proteomics, Volume 73, Issue 2, 1 December 2009, Pages 252-266

doi:10.1016/j.jprot.2009.09.009

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

KTH results – Slide P had a 100 times lower background than Corning Epoxide

1:500

1:2000

1:8000

Biotin-labeled seraEpoxy

CorningSlide P Schott

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

KTH Print buffer contained Trehalose & Pluronic F127

5% 5%APOH

APOA4

Trehalose in the plate

Trehalose before printing

data obtained with non contact printer

Nano-Plotter 2.0/E

COMPANY PRESENTATION Product Group Nexterion

Assay conditions not described

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Storage at 4°C for 4 months (KTH data)

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Effect of environmental conditions on NEXTERION Slide P performance

verschiedene "über Nacht Lagerung" von P Slides

0

5.000

10.000

15.000

20.000

25.000

30.000

35.000

Tecan Scanner 532nm 140PMT 40µm scan resolution

Inte

nsi

tät

o/n 18°C/45%LF Q alter HP

o/n RT/45%LF Q alter HP

o/n RT/Argon Q alter HP

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Is NHS hydrolysis an issue during long print runs?

Acceptable slide performance after 2 days exposure to 30 – 70 % relative humidity prior to printing

Intensität in Anhängigkeit von Luftfeuchte und Zeit der P-Slides

15000

17000

19000

21000

23000

25000

27000

29000

31000

0 1 2 3 4 5 6 7 8 9

Zeit in Tagen

Ine

tbsi

tät

35% LF

48% LF

70% LF

Ref. Argon

COMPANY PRESENTATION PRODUCT GROUP NEXTERION

Conclusion

NEXTERION Slide P has the best S/N ratios of all the

SCHOTT slide coatings for antibody arrays

Very low non specific binding, and probably lowest of any

commercially available microarray slide on the market