A rapid method for relative quantification of N-glycans from a therapeutic monoclonal antibody during trastuzumab biosimilar development. Issue 1 (1st January 2020)

• Record Type: Journal Article
• Authors: Segu, Zaneer; Stone, Todd; Berdugo, Claudia; Roberts, Anthony; Doud, Emma; Li, Yunsong
• Is Part Of: MAbs. Volume 12:Issue 1(2020)
• Publication Date: 2020-01-01
• Subjects: N-glycans -- glycoprotein -- monoclonal antibody (mAb) -- IgG -- 2-aminobenzamide (2-AB) label -- biosimilar -- herceptin antibody -- trastuzumab -- Fc-containing proteins -- protein A (PA) -- InstantPC labeling agent (IPC) -- hydrophilic interaction liquid chromatography (HILIC); Monoclonal antibodies -- Therapeutic use -- Periodicals; Monoclonal antibodies -- Periodicals; Antibodies, Monoclonal -- Periodicals; 616.0798
• DOI: 10.1080/19420862.2020.1750794
• Languages: English
• ISSNs: 1942-0862

Abstract

ABSTRACT: Glycosylation is a common post-translational modification and critical quality attribute that can modulate the efficacy of therapeutic proteins. In the production of monoclonal antibodies (mAbs), quantifying the glycoform profile is a vital characterization step. Traditional glycan analysis is time consuming and involves steps at extreme temperature or pH, which may alter glycans. Here, we describe a rapid method for glycan analysis in which glycans are released from mAb samples that are bound to protein A columns. Since host cell proteins, which may also contain glycans, were already removed, this step enables analysis of cell culture products. Glycans released from the mAb samples are then derivatized with InstantPC™ labeling agent and analyzed by HILIC-FLD-MS. To illustrate the method, the glycan profiles of six trastuzumab (Herceptin®) antibody lots and four biosimilar developmental lots were analyzed. The results derived from our novel method, which takes less than 90 min, are compared with those from a typical glycan preparation approach.