《自然》子刊：完全利用细菌生产抗体的新方法

生物谷

    最近，德克萨斯大学研究人员研制出一种完全利用细菌大规模生产抗体的方法。

    此方法是George  Georgiou博士率领的研究小组在一种鉴别新药方法的基础上加以改进得到的。之前，药物公司利用这种方法生产治疗风湿性关节炎、癌症等疾病的抗体，既费时又费力，新方法不仅克服了这项缺陷而且生产出的抗体副作用很小。

    细菌的培养成本很低，因此，无毒的E.coli早已被用于工业生产抗体。不幸的是生产抗体的方法都要构建细菌表达系统和哺乳动物细胞表达系统，费时费力，而且用哺乳动物细胞法规模生产抗体成本很高。

    Georgiou博士的E－克隆抗体法使大肠杆菌产生的抗体被锁定在细胞膜，通过向编码抗体的基因引入小“错误”，增强抗体攻击疾病蛋白的能力。抗体和疾病蛋白之间的这种相互作用不仅有效地缩短了治疗时间，而且阻滞了蛋白对肌体的危害。

    为了检测这个细菌为唯一成分的系统，第一作者Yariv  Mazor设计出一种抗炭疽热毒素PA的抗体，然后通过APEx方法（由Georgiou实验室和Iverson实验室合作完成）证实细菌产生的抗体与PA的结合力很强。研究人员筛选出这些细菌并大量培养，进一步精炼大规模生产治疗性抗体的步骤。研究结果刊登于4月15日《Nature  Biotechnology》。

原始出处:

Nature Biotechnology，Published online: 15 April 2007; | doi:10.1038/nbt1296

Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli

Yariv Mazor1, 3, Thomas Van Blarcom1, 3, Robert Mabry1, 2, Brent L Iverson1, 2 & George Georgiou1, 3, 4, 5

1 Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.

2 Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas 78712, USA.

3 Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712, USA.

4 Department of Biomedical Engineering, University of Texas at Austin, Austin, Texas 78712, USA.

5 Section of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, Texas 78712, USA.

Correspondence should be addressed to George Georgiou gg@che.utexas.edu

Abstract

We describe facile isolation of full-length IgG antibodies from combinatorial libraries expressed in E. coli. Full-length heavy and light chains are secreted into the periplasm, where they assemble into aglycosylated IgGs that are captured by an Fc-binding protein that is tethered to the inner membrane. After permeabilizing the outer membrane, spheroplast clones expressing so-called E-clonal antibodies, which specifically recognize fluorescently labeled antigen, are selected using flow cytometry. Screening of a library constructed from an immunized animal yielded several antibodies with nanomolar affinities toward the protective antigen of Bacillus anthracis.

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