Anushikha Thakur1, Rekha Nagpal1, Avik Kumar Ghosh2, Deepak Gadamshetty3, Sirisha Nagapattinam4, Malini Subbarao1, Shreshtha Rakshit1, Sneha Padiyar1, Suma Sreenivas1, Nagaraja Govindappa1, Harish V Pai1, Ramakrishnan Melarkode Subbaraman5. 1. Research and Development (RND), Biocon Biologics Limited, Biocon Park, SEZ, Plot No. 2&3,Phase-IV, Bommasandra Industrial Estate, Bommasandra-Jigani Link Road, Bangalore, 560099, India. 2. Alvotech Hf, Saemundargata 15-19, 101, Reykjavik, Iceland. 3. Divis Laboratories Limited, Chotuppal, Hyderabad, 508252, India. 4. Sunquest Information Systems, Bangalore, 560095, India. 5. Research and Development (RND), Biocon Biologics Limited, Biocon Park, SEZ, Plot No. 2&3,Phase-IV, Bommasandra Industrial Estate, Bommasandra-Jigani Link Road, Bangalore, 560099, India. ramakrishnan.melarkode@biocon.com.
Abstract
Sequence variants (SV) in protein bio therapeutics can be categorized as unwanted impurities and may raise serious concerns in efficacy and safety of the product. Early detection of specific sequence modifications, that can result in altered physicochemical and or biological properties, is therefore desirable in product manufacturing. Because of their low abundance, and finite resolving power of conventional analytical techniques, they are often overlooked in early drug development. Here, we present a case study where trace amount of a sequence variant is identified in a monoclonal antibody (mAb) based therapeutic protein by LC-MS/MS and the structural and functional features of the SV containing mAb is assessed using appropriate analytical techniques. Further, a very sensitive selected reaction monitoring (SRM) technique is developed to quantify the SV which revealed both prominent and inconspicuous nature of the variant in process chromatography. We present the extensive characterization of a sequence variant in protein biopharmaceutical and first report on control of sequence variants to < 0.05% in final drug product by utilizing SRM based mass spectrometry method during the purification steps.
Sequence variants (SV) in protein bio therapeutics can be categorized as unwanted impurities and may raise serious concerns in efficacy and safety of the product. Early detection of specific sequence modifications, that can result in altered physicochemical and or biological properties, is therefore desirable in product manufacturing. Because of their low abundance, and finite resolving power of conventional analytical techniques, they are often overlooked in early drug development. Here, we present a case study where trace amount of a sequence variant is identified in a monoclonal antibody (mAb) based therapeutic protein by LC-MS/MS and the structural and functional features of the SV containing mAb is assessed using appropriate analytical techniques. Further, a very sensitive selected reaction monitoring (SRM) technique is developed to quantify the SV whn class="Disease">ich revealed both prominent and inconspicuous nature of the variant in process chromatography. We present the extensive characterization of a sequence variant in protein biopharmaceutical and first report on control of sequence variants to < 0.05% in final drug product by utilizing SRM based mass spectrometry method during the purification steps.
Authors: X Christopher Yu; Oleg V Borisov; Melissa Alvarez; David A Michels; Yajun Jennifer Wang; Victor Ling Journal: Anal Chem Date: 2009-11-15 Impact factor: 6.986
Authors: Lauren Feeney; Veronica Carvalhal; X Christopher Yu; Betty Chan; David A Michels; Yajun Jennifer Wang; Amy Shen; Jan Ressl; Brendon Dusel; Michael W Laird Journal: Biotechnol Bioeng Date: 2013-02-15 Impact factor: 4.530
Authors: Jun Lin; Mengyu Xie; Dan Liu; Zhen Gao; Xiaoyan Zhao; Hongxia Ma; Sheng Ding; Shu Mei Li; Song Li; Yanling Liu; Fang Zhou; Hao Hu; Tao Chen; He Chen; Min Xie; Bo Yang; Jun Cheng; Mingjun Ma; Yanyang Nan; Dianwen Ju Journal: Front Chem Date: 2022-09-20 Impact factor: 5.545