PT  - JOURNAL ARTICLE
AU  - Ming Yao
AU  - Bingming Chen
AU  - Weiping Zhao
AU  - John T. Mehl
AU  - Lingjun Li
AU  - Mingshe Zhu
TI  - LC-MS Differential Analysis for Fast and Sensitive Determination of Biotransformation of Therapeutic Proteins
AID  - 10.1124/dmd.117.077792
DP  - 2018 Apr 01
TA  - Drug Metabolism and Disposition
PG  - 451--457
VI  - 46
IP  - 4
4099  - http://dmd.aspetjournals.org/content/46/4/451.short
4100  - http://dmd.aspetjournals.org/content/46/4/451.full
SO  - Drug Metab Dispos2018 Apr 01; 46
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.