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 Jan 01
TA  - Drug Metabolism and Disposition
PG  - dmd.117.077792
4099  - http://dmd.aspetjournals.org/content/early/2018/01/31/dmd.117.077792.short
4100  - http://dmd.aspetjournals.org/content/early/2018/01/31/dmd.117.077792.full
AB  - Therapeutic biologics have become a fast-growing segment within the pharmaceutical industry during the past three 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 modification. 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 thirty-eight peptides were significantly changed. Moreover, while all modifications were screened and detected, only the ones related to the treatment process were revealed by differential analysis. Other modifications that co-exist 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 biologics drug discovery and development.