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生成用于检测淋巴丝虫重组抗原 BmR1 和 BmSXP 的人源 scFv-IgG1Fc 抗体。

Generation of human scFv-IgG1Fc antibodies for detection of lymphatic filarial recombinant antigens, BmR1 and BmSXP.

机构信息

Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden, Penang, Malaysia.

出版信息

Biotechnol Appl Biochem. 2022 Feb;69(1):70-76. doi: 10.1002/bab.2082. Epub 2020 Dec 8.

Abstract

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.

摘要

淋巴丝虫病是一种被忽视的寄生虫病,影响着热带和亚热带国家的数百万人,由班氏丝虫和马来丝虫引起。在需要快速检测的情况下,在欠发达和偏远地区进行检测,以覆盖受感染地区,这对于消除该疾病作为公共卫生问题至关重要。有一些基于抗原或抗体检测的商业化快速检测方法,但前者仅检测到由班氏丝虫引起的感染,并且与非淋巴丝虫发生交叉反应,而抗体检测可能会提供以前感染的阳性结果。在这里,我们报告了三种不同的基于单链抗体 (scFv) 的重组免疫球蛋白 γ (IgG1) 抗体的产生,这些 scFv 是通过人抗体噬菌体展示技术先前产生的,即抗 BmR1 克隆 4、抗 BmXSP 克隆 5B 和抗 BmXSP 克隆 2H2。scFv 序列被克隆到 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够与各自的靶标结合。Fc 与 scFv 的缀合诱导结合物稳定性,并为探针和信号分子提供多个标记位点,可用于快速检测。

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