As title describes, we're trying to mutate an amino acid of antibody Fc to cysteine, followed by TCEP to further drug conjugation, one of my colleague had raise the concern of purified TCEP treated antibody might be affected by the tris that had been used to store the protein. Is it possible? and if yes, what might be the reason?
John Carter
No. Tris is a problem with succinimide conjugation, but it won't affect maleimide conjugation. You may expect TCEP to reduce most or all Cys in the Ab, not only your newly introduced residue. So you may observe multiple coupling, especially in the hinge region. Degas and avoid aeration, as Cys undergoes facile air oxidation.
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