Generation of human scFv–IgG1Fc antibodies for detection of lymphatic filarial recombinant antigens, BmR1 and BmSXP,Biotechnology and Applied Biochemistry

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Generation of human scFv–IgG1Fc antibodies for detection of lymphatic filarial recombinant antigens, BmR1 and BmSXP
Biotechnology and Applied Biochemistry ( IF 3.2 ) Pub Date : 2020-11-30 , DOI: 10.1002/bab.2082
Ahmad Ismail Khaled Abdo ₁ , Ying Yuan Ngoh ₁ , Min Han Lew ₁ , Sylvia Annabel Dass ₁ , Anizah Rahumatullah ₁ , Rahmah Noordin ₁ , Gee Jun Tye ₁

Affiliation

Lymphatic filariasis is a neglected parasitic disease that affects millions in tropical and subtropical countries and is caused by Wuchereria and Brugia species. Specific and sensitive detection methods are essential in mapping infected areas where rapid tests are needed to cover underdeveloped and remote regions, which facilitates eliminating the disease as a public health problem. A few commercialized rapid tests based on antigen or antibody detection are available, but the former only detects infection by Wuchereria species and cross-reacts with nonlymphatic filaria, whereas antibody detection might provide positive results of previous infection. Here, we report the production of three different recombinant immunoglobulin gamma (IgG)1 antibodies based on scFvs previously generated via human antibody phage display technology, that is, anti-BmR1 clone 4, anti-BmXSP clone 5B, and anti-BmXSP clone 2H2. The scFv sequences were cloned into a pCMV-IgG1 vector, then transfected into a HEK293F cell line. The generated antibodies were found to be able to bind to their respective targets even at relatively low concentration. Conjugation of Fc to scFv induces binder stability and provides multiple labeling sites for probes and signaling molecules that can be used in rapid tests.

中文翻译：

产生用于检测淋巴丝虫重组抗原、BmR1 和 BmSXP 的人 scFv-IgG1Fc 抗体

淋巴丝虫病是一种被忽视的寄生虫病，在热带和亚热带国家影响数百万人，由Wuchereria和Brugia物种引起。具体和敏感的检测方法对于绘制需要快速检测以覆盖欠发达和偏远地区的受感染区域至关重要，这有助于消除该疾病作为公共卫生问题。有一些基于抗原或抗体检测的商业化快速检测，但前者仅检测乌彻氏菌感染种并与非淋巴丝虫发生交叉反应，而抗体检测可能提供先前感染的阳性结果。在这里，我们报告了基于先前通过人抗体噬菌体展示技术产生的 scFv 的三种不同重组免疫球蛋白 γ (IgG)1 抗体的生产，即抗 BmR1 克隆 4、抗 BmXSP 克隆 5B 和抗 BmXSP 克隆 2H2 . 将 scFv 序列克隆到 pCMV-IgG1 载体中，然后转染到 HEK293F 细胞系中。发现生成的抗体即使在相对较低的浓度下也能与它们各自的靶标结合。Fc 与 scFv 的结合可诱导结合剂稳定性，并为可用于快速测试的探针和信号分子提供多个标记位点。

更新日期：2020-11-30

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