Affinité SPR Workflow

workflow2
Figure 1. Affinité SPR workflow

There is one offline SAM preparation step and six online injections steps to build SPR sensor layer-by-layer prior probing the sample (blue region). The real-time layer building offers plenty checkpoints to ensure the quality of assay. Chips can be prepared and stored ahead of time at various stages of the process for faster testing.

 

Process Step Typical Material Step Time Storage Time / Conditions
c1  Self-Assembled Monolayer Alkyl Thiol (-COOH terminated), Peptide, Dextran Overnight (8h) Months / Dry, inert environment
c2  Activation EDC - NHS, Cysteamine - Glutaraldehyde 2-10 min - - -
c3  Immobilization Peptide (antigen, antibody, membrane) or DNA* 15-30 min - - -/- - -
c4  Blocking Ethanolamine, Mercaptohexanol, BSA 15-30 min Weeks/PBS, 4°C
c5  Detection Buffer, Body fluid, Water, Juice, Milk, etc. 2 min - hours - - -/- - -
c6  Regeneration Urea, Phosphoric Acid, NaOH, NaCl, Tween 20 10 - 20 seconds Hours - Days/PBS, 4°C or RT

*Thiolated DNA oligo immobilization does not require the initial two steps