I performed passive adsorption of protein on my PS microspheres, but there is some minor visible clumping in the final solution. I had performed all steps as per a previously used protocol that didn't pose any problem. What can I do to fix this issue without losing any of the bound antibody (anti-PSA) or blocking agent (BSA).
One method I have read is through dialysis, but I do not want to remove any protein from the surface since my microspheres contain much larger amount of BSA than the antibody, and rebinding may effectively reduce antibody concentration on the spheres.
If all is the same as before then I would worry about whether your coating antigen is already complexed. Even a small amount of complex formation in your protein could produce the results you describe. You could try a quick ultra spin, or filtration to remove complexes before coating. Be sure you do not alter the concentration too much if you do this.
Thanks for your response. I tried to vortex the solution and it worked to remove most of the bigger clumps. The remaining smaller clumps dissolved on their own over time. I guessed it may have been because of the centrifuge step.
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