Overview of SCHOTT’s protein micro-array slide surfaces
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Choosing the best slide coating chemistry for your protein micro-array application. Poster presented at the Advances in Microarray Technology 2008 conference in Barcelona, Spain.
![Substrate Formats ,[object Object],[object Object],SCHOTT’s Protein Microarray Slides: Choosing the right coating Dietmar Knoll, Corinna Schwung, Alistair Rees and Rüdiger Dietrich SCHOTT Technical Glass Solutions GmbH., Otto-Schott-Strasse 13, 07745 Jena, Germany +49 3641 681 4066 NEXTERION ® protein microarray surfaces MTP-96 well SBS microplate format 16-pad slide Standard 25 x 75 mm slide Customized 8-well slide (customized) 2-pad slide The lower number of protein species to be printed allows smaller array sizes. This allows lower assay volumes and finally cost/time savings due to multiplexing. Conventional 2D surfaces (<10nm) Metallic surfaces 3-D surfaces Epoxy, Aldehyde-silane Gold coatings Protein Microarray Substrates 3-D thick film coatings (0.1–200µm) 3-D thin film coatings (10–100nm) Nitrocellulose, Agarose coatings Polymer and hydrogel type Conventional 2-D Substrates 3-D Thick Film Coatings-Nitrocellulose SEM image of NEXTERION ® Slide NC Schott Nexterion ® Slide NC Diagram showing increased capacity due to pore structure Examples 1. NC (NEXTERION ® Slide NC-N and -C) 2. Agarose Remarks 1. Nitrocellulose is widely used surface for proteins 2. High print buffer tolerance (e.g. lysate detergents) 3. Stabilization of protein structure (long shelf life) 4. Can have high background 5. Opaque NC coating not compatible some detection methods 3-D Thin Film Coatings Binding Capacity and Signal Intensities ,[object Object],[object Object],[object Object],Background ,[object Object],[object Object],[object Object],Signal / Background ratio Benchmark Test Results Summary Printed probe: rabbit anti goat Ab (1,2mg/ml) – target: IgG Goat ,[object Object],[object Object],[object Object],S/B ratio Binding capacity and signal intensity. (0,25) Signal/Background ratio (0,25) Probe target linearity (0,2) Sensitivity (0,15) Print buffer tolerance (0,15) Printed slide storage conditions Slide H +++ ++++ +++ ++++ ++ -20 °C Slide P +++ ++++ +++ ++++ ++ -20 °C Slide NC ++++ ++ ++++ ++ ++++ +20 °C Slide E +++ +++ +++ +++ +++ - 20 °C Slide AL ++ ++ ++ ++ ++ -20 °C SCHOTT Nexterion CH 2 CH O l CH 2 CH O l CH 2 CH O l epoxy groups Immobilization •• O NH 2 OH NH P P ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Cross links Reactive group Polymer - selected to provide lowest unspecific binding and to stabilize protein structure ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Data have been normalized to allow data comparison Printed probe: rabbit anti goat Ab – target: IgG Goat (3,125µg/ml) signal intensity colours refer to antibody concentration printed Slide H Slide P Slide E Slide AL Slide NC 0 100000 200000 300000 400000 500000 600000 700000 800000 900000 1000000 02 RaG 0,01mg/ml 3125 03 RaG 0,025mg/ml 3125 04 RaG 0,1mg/ml 3125 05 RaG 0,3mg/ml 3125 06 RaG 0,6mg/ml 3125 07 RaG 1,2mg/ml 3125 Slide H Slide P Slide E Slide AL Slide NC signal intensity 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000 02 RaG 0,01mg/ml 3125 03 RaG 0,025mg/ml 3125 04 RaG 0,1mg/ml 3125 05 RaG 0,3mg/ml 3125 06 RaG 0,6mg/ml 3125 07 RaG 1,2mg/ml 3125 colours refer to antibody concentration printed 0 Slide H Slide P Slide E Slide AL Slide NC signal intensity colours refer to target concentration (IgG goat) 0 100 200 300 400 500 600 3125 625 125 25 5 1 signal intensity colours refer to target concentration (IgG goat) 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 intensity 3125 625 125 25 5 1 Slide H Slide P Slide E Slide AL Slide NC Slide H Slide P Slide E Slide AL Slide NC Printed probe: anti osteopontin Ab – target: Osteopontin S/B ratio Printed protein concentration in mg/ml colours refer to target con- centration (osteopontin) 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 0,03125 0,0625 0,125 0,25 0,5 1 0,03125 0,0625 0,125 0,25 0,5 1 0,03125 0,0625 0,125 0,25 0,5 1 0,03125 0,0625 0,125 0,25 0,5 1 0,03125 0,0625 0,125 0,25 0,5 1 3125 ng/ml 625 ng/ml 125 ng/ml 25 ng/ml 5 ng/ml 1 ng/ml Printed protein concentration in mg/ml Slide H Slide P Slide E Slide AL Slide NC](https://image.slidesharecdn.com/posterschottamt-100908083506-phpapp02/85/Overview-of-SCHOTT-s-protein-micro-array-slide-surfaces-1-320.jpg)
