抗精子抗体的平台特异性 Fc N-糖谱分析

IF 3 Q3 IMMUNOLOGY

Antibodies Pub Date : 2024-03-06 DOI:10.3390/antib13010017

Ellena Nador, Chaoshuang Xia, Philip J. Santangelo, Kevin J. Whaley, Catherine E. Costello, Deborah J. Anderson

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引用次数: 0

摘要

IgG Fc N-糖基化是发挥效应功能的必要条件，也是质量控制的重要组成部分。抗体生产平台的选择有可能极大地影响抗体的 Fc 糖，从而改变其活性和临床特征。人类避孕抗体 (HCA) 是一种 IgG1 抗精子单克隆抗体 (mAb)，目前正作为一种新型非激素避孕药进行临床开发。其开发工作的一部分是选择合适的表达平台来生产用于女性生殖道的 HCA。在这里，我们比较了两种新型 mAb 生产平台（即转基因烟草植物（Nicotiana benthamiana; HCA-N）和 mRNA 介导的人阴道细胞表达（HCAmRNA））生产的 HCA 的 Fc 糖基化。使用质谱法测定了两种 HCA 产品的 Fc N-糖谱。结果显示，两种 HCA 产物在位点占有率、聚糖类型和糖型分布上存在重大差异。为了解决这些差异如何影响 Fc 功能的问题，我们进行了抗体依赖性细胞吞噬（ADCP）试验。在 HCA-N 存在的情况下，精子吞噬水平明显低于 HCAmRNA。这项研究提供了两种 HCA 生产平台生产功能不同的 HCA 的证据；这些信息可能有助于为 HCA 临床开发和一般 mAbs 选择最佳平台。

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Platform-Specific Fc N-Glycan Profiles of an Antisperm Antibody

IgG Fc N-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 (Nicotiana benthamiana; HCA-N) and mRNA-mediated expression in human vaginal cells (HCAmRNA). The Fc N-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 HCAmRNA. 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.

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来源期刊

Antibodies IMMUNOLOGY-

CiteScore

7.10

自引率

6.40%

发文量

审稿时长

11 weeks

期刊介绍： Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.