We use 990 ul of antibody solution and then add 0.4mg EDC and 1.1.mg sulfo-NHS (in MES) and react for 15 minutes at room temperature, before adding it to the amine surface for immobilization. Now when we are looking to scale this process to coat tens and hundreds of units for clinical studies, we are unable to follow this process due to manual labour required for each step, and are trying to automate it at lab scale.
Firstly I wanted to know how long can this solution provide the same performance at Room temperature? Secondly, how can we increase the shelf life for this to longer period (at least 30-45 minutes) to ensure high precision and repeatability? We don't have budget for a fluid injection and dosing system for automation.
This is a very risky approach, and I would strongly recommend that you reverse the chemistry and conjugate to a carboxyl surface. By adding EDC and sulfo-NHS to an antibody, you are activating carboxyls on a molecule that already has 70-100 lysines. Thus, crosslinking of the Ab is going to be a major issue, and there is no possibility of getting consistent performance after storage for different times. A preferred approach is to activate a carboxyl surface and add sNHS groups, and then add the antibody last. In this way, the antibody cannot react with itself and you have a surface that is reasonably stable over time, as long as the pH is not too high.
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