期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 130, 期 3, 页码 327-333出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2020.04.009
关键词
Monoclonal antibody; Critical quality attribute; N-linked glycosylation; Sample preparation; High-performance liquid chromatography; High resolution profile
资金
- NSF [1706731, 1624718]
- Directorate For Engineering
- Div Of Chem, Bioeng, Env, & Transp Sys [1706731] Funding Source: National Science Foundation
- Div Of Industrial Innovation & Partnersh
- Directorate For Engineering [1624718] Funding Source: National Science Foundation
N-linked glycosylation is a post-translational modification that occurs on many proteins during biosynthesis. The profile of different glycans on the protein is a critical quality attribute of some recombinant biopharmaceutical proteins including monoclonal antibodies (mAbs). Methods for profiling glycan should be robust, fast, and sensitive. Isolating glycans from proteins and tagging a label on glycans is the most commonly used technique for glycan profiling. Currently, existing protocols for sample preparation can be complicated, time-consuming, and expensive, which can limit the wide adaptation of glycan profiling methods. As a further barrier to use, an expensive ultra-high-pressure liquid chromatography (UHPLC) system is frequently required for the profile. In this article, a low cost and easily-used workflow of sample preparation is coupled with a standard high-performance liquid chromatography (HPLC) systemto achieve comparable results to UHPLC. The number of steps required in the protocol and the time, as well as the cost associated with the sample preparation, is significantly reduced, while maintaining robust analytical performance. We describe the creation and validation of a human serum IgG glycan library to be used as the calibration standard, and successful profiling of glycoforms from a variety of mAbs. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.
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