RT Journal Article
SR Electronic
T1 LC-MS Differential Analysis for Fast and Sensitive Determination of Biotransformation of Therapeutic Proteins
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 451
OP 457
DO 10.1124/dmd.117.077792
VO 46
IS 4
A1 Ming Yao
A1 Bingming Chen
A1 Weiping Zhao
A1 John T. Mehl
A1 Lingjun Li
A1 Mingshe Zhu
YR 2018
UL http://dmd.aspetjournals.org/content/46/4/451.abstract
AB Therapeutic biologics have become a fast-growing segment within the pharmaceutical industry during the past 3 decades. Although the metabolism of biologics is more predictable than small molecule drugs, biotransformation can significantly affect the activity of biologics. Unfortunately, there are only a limited number of published studies on the biotransformation of biologics, most of which are focused on one or a few types of modifications. In this study, an untargeted LC-MS–based differential analysis approach was developed to rapidly and precisely determine the universal biotransformation profile of biologics with the assistance of bioinformatic tools. A human monoclonal antibody (mAb) was treated with t-butyl hydroperoxide and compared with control mAb using a bottom-up proteomics approach. Thirty-seven types of post-translational modifications were identified, and 38 peptides were significantly changed. Moreover, although all modifications were screened and detected, only the ones related to the treatment process were revealed by differential analysis. Other modifications that coexist in both groups were filtered out. This novel analytical strategy can be effectively applied to study biotransformation-mediated protein modifications, which will streamline the process of biologic drug discovery and development.