I'll add some chemical like ammonium acetate and sodium borohydrate for my experiment. In this mixture, does the antibody stay stable? How can I check that the antibody stays without any cleavage or hydrolysis? Is there any other method to do it without ELISA?
If you are only worried about cleavage or hydrolysis, run SDS-PAGE. If you are worried about the ability of the antibody to recognize its epitope, do a quantitative immunological assay, such as ELISA. You could also check for aggregation using dynamic light scattering.
Hi Jun, are you working with monoclonal or polyclonal antibodies?
Monoclonals (MoAbs) are notorious for having different stabilities. These properties will have to be determined empirically for each one.
There are no general assays that I know of, besides ELISA, that will give you information on MoAb stability.
Well, if you rule out the cleavage and hydrolysis by PAGE and you are still not sure about stability being influenced, you could go for the thermofluor assay (often used as pre-screening for crystallography to determine the most stable buffer conditions for protein). You would only need a suitable dye (Sypro Orange) and qPCR cycler. It is super fast and super simple (you can check on wikipedia), requires small amounts of protein and would help to determine if the stability is impaired after addition of your special chemical, compared to original buffer.
If you have plenty of your antibody and don't care about the amount used for screening, there are other methods, which can be used: like differential scanning calorimetry..
From experience (conjugating antibodies to HRP), AB stability should not be affected much. However, characterization by ThermofluorAssay, as Veronika suggested, might be an interesting option.
As NaBH4 is alkaline, make sure you have a suitable buffer capacity for your experiment.
If you have access to circular dichroism, that would show you the unfolding, and any change in the secondary structure of the antibody. Antibodies have distinctive absorption bands on CD.
You could perform nanoDSF to test conformational and colloidal stability of your antibody. nanoDSF works in any buffer, since no dye and no labeling is required. And you can easily do forced-degradation tests.
http://www.nanotemper-technologies.com/get-it-all/application-notes/nanodsf/
You also might call your antibody vendor. Their staff scientists may know about stability in these particular compounds.
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