Free sulfhydryl measurement as an indicator of antibody stability.
Monoclonal antibodies are a major subclass of biopharmaceuticals. They are structurally different from other biopharmaceuticals in size and quaternary structure. Here we demonstrate a correlation between chemical stability of antibodies and thermal stability. We show that overall thermal protein stability can be predicted based on the measurement of free sulfhydryl (-SH) content on applying mildly denaturing conditions. We propose that this method can be adapted to a high-throughput screening format and used either as an absolute measure of thermal stability or for ranking a panel of possible variants.
Lacy
Eilyn R
ER
Protein Engineering Department, Centocor R&D, Inc., Radnor, PA 19087, USA.
Baker
Margaret
M
Brigham-Burke
Michael
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D002138:Calibration; D002417:Cattle; D005453:Fluorescence; D006801:Humans; D011489:Protein Denaturation; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000737:chemistry*; D044366:Transition Temperature
D000911:Antibodies, Monoclonal; D013438:Sulfhydryl Compounds