Application of a 2A-peptide system for polycistronic human IgG production in Leishmania tarentolae

IF 3.6 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods Pub Date : 2026-02-01 Epub Date: 2025-12-15 DOI:10.1016/j.ymeth.2025.12.005

Jing Yi Lai , Kae Yi Tan , Choo Hock Tan , Yee Siew Choong , Theam Soon Lim

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引用次数: 0

Abstract

Monoclonal antibodies are commonly expressed using mammalian cell lines due to their required post-translational modifications. However, this process can become costly and requires investment in specialized equipment and facilities. The use of eukaryotic cell Leishmania tarentolae offers a cost-effective alternative while still allowing N-linked glycosylation to occur. The focus of this study explores the use of L. tarentolae as a platform for expressing a monoclonal IgG antibody against the long-chain neurotoxin (LNTX) from the venom of the Monocled Cobra, Naja kaouthia. The monoclonal antibody was isolated from a human naïve phage display library and expressed as single-chain fragment variable (scFv) in Escherichia coli with good specificity against the LNTX of Naja kaouthia. The scFv was then converted to the scFv-Fc format and full-length IgG for expression in L. tarentolae. The IgG expression was achieved using a 2A peptide-based bicistronic vector system with good expression yield. The expressed IgG demonstrated homogeneity in size and minimal degradation. Therefore, L. tarentolae can be considered a possible alternative platform for the expression of human scFv-Fc and IgG antibodies.

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2a肽体系在产人多顺反性IgG中的应用。

单克隆抗体通常在哺乳动物细胞系中表达，因为它们需要翻译后修饰。然而，这个过程可能会变得昂贵，需要投资专门的设备和设施。使用真核细胞利什曼绦虫提供了一种具有成本效益的替代方案，同时仍然允许n -链糖基化发生。本研究的重点是探索利用L. tarentolae作为表达单克隆IgG抗体的平台，抗来自单眼眼镜蛇Naja kaouthia毒液的长链神经毒素（LNTX）。该单克隆抗体从人naïve噬菌体展示文库中分离得到，在大肠杆菌中以单链片段变量（single-chain fragment variable, scFv）的形式表达，具有较好的特异性。然后将scFv转化为scFv- fc格式和全长IgG，在猪乳杆菌中表达。IgG的表达采用基于2A肽的双链载体系统，表达量高。表达的IgG大小均匀，降解程度最低。因此，可以将链乳杆菌视为表达人scFv-Fc和IgG抗体的可能替代平台。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods 生物-生化研究方法

CiteScore

9.80

自引率

2.10%

发文量

222

审稿时长

11.3 weeks

期刊介绍： Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.