Nador Ellena, Xia Chaoshuang, Santangelo Philip J, Whaley Kevin J, Costello Catherine E, Anderson Deborah J
Department of Medicine, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA.
Center for Biomedical Mass Spectrometry, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA.
Antibodies (Basel). 2024 Mar 6;13(1):17. doi: 10.3390/antib13010017.
IgG Fc -glycosylation is necessary for effector functions and is an important component of quality control. The choice of antibody manufacturing platform has the potential to significantly influence the Fc glycans of an antibody and consequently alter their activity and clinical profile. The Human Contraception Antibody (HCA) is an IgG1 antisperm monoclonal antibody (mAb) currently in clinical development as a novel, non-hormonal contraceptive. Part of its development is selecting a suitable expression platform to manufacture HCA for use in the female reproductive tract. Here, we compared the Fc glycosylation of HCA produced in two novel mAb manufacturing platforms, namely transgenic tobacco plants (; HCA-N) and mRNA-mediated expression in human vaginal cells (HCA). The Fc -glycan profiles of the two HCA products were determined using mass spectrometry. Major differences in site occupancy, glycan types, and glycoform distributions were revealed. To address how these differences affect Fc function, antibody-dependent cellular phagocytosis (ADCP) assays were performed. The level of sperm phagocytosis was significantly lower in the presence of HCA-N than HCA. This study provides evidence that the two HCA manufacturing platforms produce functionally distinct HCAs; this information could be useful for the selection of an optimal platform for HCA clinical development and for mAbs in general.
IgG Fc糖基化对于效应功能是必需的，并且是质量控制的重要组成部分。抗体生产平台的选择有可能显著影响抗体的Fc聚糖，从而改变其活性和临床特征。人避孕抗体（HCA）是一种IgG1抗精子单克隆抗体（mAb），目前正处于临床开发阶段，作为一种新型的非激素避孕药。其部分研发工作是选择合适的表达平台来生产用于女性生殖道的HCA。在此，我们比较了在两种新型单克隆抗体制备平台中产生的HCA的Fc糖基化情况，这两种平台分别是转基因烟草植株（；HCA-N）和人阴道细胞中的mRNA介导表达（HCA）。使用质谱法测定了两种HCA产品的Fc聚糖谱。结果显示在位点占有率、聚糖类型和糖型分布方面存在主要差异。为了探究这些差异如何影响Fc功能，进行了抗体依赖性细胞吞噬（ADCP）试验。在HCA-N存在的情况下，精子吞噬水平显著低于HCA。本研究提供了证据，证明这两种HCA生产平台产生功能不同的HCA；这些信息对于为HCA临床开发选择最佳平台以及一般单克隆抗体而言可能是有用的。
