Some of the most accessible hinge SS or maybe even the light-heavy chain SS may be attacked, however, this will not destroy the antibody once it is fully folded. The intra-domain SS bridges are well buried in the core of the domains, they are unlikely to get reduced under non-denaturing conditions.

Did you realize that somebody asked exactly the same question six years ago? https://www.researchgate.net/post/DTT_use_for_sputum_samples_with_ELISA.

As a careful researcher, I would do a test run on controls and a few samles with and without DTT before embarking on any large-scale screening effort, although I am pretty convinced that under non-denaturing conditions, the reaction of the buried disulfides would be too slow to cause problems, although it might depend on the specific antibody and antigen pair.

These researchers Article Effect of sputum processing with dithiothreitol on the detec...

found that:

"Median levels of IL-1ß, IL-6, IL-8, SLPI, and NE were similar in the DTT and NaCl treated samples. There was a significant reduction in median (IQR) levels of detectable TNFα (0.07 (0.00–0.47) pM v 0.90 (0.06–6.98) pM, p

Thank you Annemarie Honegger for your answer!

I have already read both the question on researchgate and the article you cited, and I am in doubt about of the different results on different cytokines.

Probably the best way to go on is to do a test with some control samples with and without DTT, as you suggested.

Exactly, you have to verify the the influence of DTT on the test results for your specific antibody/antigen pairs. Depending on your antigen, this may have more accessible disulfide bonds than the antibody variable domains, and therefore the conformation of the epitope recognized by your antibody may also be affected DTT reduction.