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应用噬菌体展示技术从体外免疫人外周血单个核细胞制备抗痤疮丙酸杆菌的人源单克隆抗体。

Generation of human monoclonal antibodies against Propionibacterium acnes by applying the phage display method to human peripheral blood mononuclear cells immunized in vitro.

机构信息

Graduate School of Bioresources and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan.

出版信息

Cytotechnology. 2008 Jun;57(2):169-75. doi: 10.1007/s10616-008-9138-z. Epub 2008 Feb 22.

Abstract

Propionibacterium acnes is a gram-positive, non-spore-forming, rod-shaped bacterium that is often detected in normal human skin flora. P. acnes has been associated with many diseases. In this study, we attempted to generate anti-P. acnes human monoclonal antibodies. A phage antibody library was first generated from human peripheral blood mononuclear cells immunized in vitro with P. acnes using the phage display method, and P. acnes-specific phage antibodies were obtained using solid phase panning. Antigen-specific variable region genes were then amplified and recombined into vectors expressing human IgG antibodies. The results indicated that the recombinant human IgG antibodies exhibited P. acnes-specific binding. This study demonstrates that the combined use of an in vitro immunization protocol and the phage display method enables the generation of human monoclonal antibodies against pathogenic bacteria and toxic antigens.

摘要

痤疮丙酸杆菌是一种革兰阳性、非孢子形成、杆状细菌,常存在于正常人体皮肤菌群中。痤疮丙酸杆菌与许多疾病有关。在本研究中,我们试图产生抗痤疮丙酸杆菌的人源单克隆抗体。首先通过噬菌体展示方法,用痤疮丙酸杆菌体外免疫人外周血单核细胞,生成噬菌体抗体库,然后通过固相淘选获得痤疮丙酸杆菌特异性噬菌体抗体。接着扩增抗原特异性可变区基因,并将其重组到表达人 IgG 抗体的载体中。结果表明,重组人 IgG 抗体表现出对痤疮丙酸杆菌的特异性结合。本研究表明,体外免疫方案和噬菌体展示方法的联合使用能够产生针对致病菌和毒性抗原的人源单克隆抗体。

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