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单个B细胞抗体制备技术及应用 被引量:14

Single B cell monoclonal antibody technologies and applications
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摘要 单克隆抗体是现代生命科学研究的重要工具,为许多领域的发展作出了不可估量的贡献。随着PCR技术和单克隆抗体技术的发展和成熟,单个B细胞抗体制备技术迅速兴起。该技术能够对单个的抗原特异性B细胞进行抗体基因的体外克隆和表达,保证了轻重链可变区的天然配对,相较于传统的抗体制备技术具有效率高、全人源、基因多样性更丰富等优势。单个B细胞抗体制备技术已成为制备全人抗体的热门方法,同时也促进了包括抗体发生成熟、疫苗保护机制、疫苗开发、肿瘤及自身免疫疾病等免疫学相关研究。文中就单个B细胞抗体制备技术的过程及应用作简要综述。 Monoclonal antibodies (mAbs) contribute a lot to the development of numerous fields in life science as a pivotal tool in modern biological research. Development of the PCR methods and maturation of antibody production have made it possible to generate mAbs from single human B cells by single cell RT-PCR with successional cloning and expression in vitro. Compared to traditional monoclonal antibody technologies, single B cell technologies require relatively fewer cells, which are highly efficient in obtaining specific mAbs in a rapid way with preservation of the natural heavy andlight chain pairing. With so many advantages, single B cell technologies have been proved to be an attractive approach for retrieval of naive and antigen-experienced antibody repertoires generated in vivo, design of rationale structure-based vaccine, evaluation and development of basic B cell biology concepts in health and autoimmunity, and prevention of infectious diseases by passive immunization and therapy for disorders. Accordingly, this review introduced recent progresses in the single B cell technologies for generating monoclonal antibodies and applications.
出处 《生物工程学报》 CAS CSCD 北大核心 2012年第6期651-660,共10页 Chinese Journal of Biotechnology
基金 国家自然科学基金(No.81172980) 国家杰出青年科学基金(No.81025018)资助~~
关键词 单克隆抗体 单个B细胞 免疫球蛋白基因 逆转录PCR monoclonal antibody, single B cell, immunoglobulin gene, RT-PCR
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  • 1Gilbert AE, Karagiannis P, Dodev T, et al. Monitoring the systemic human memory B cell compartment of melanoma patients for anti-tumor IgG antibodies. PLoS One, 2011, 6(4): e19330.
  • 2Morris L, Chen X, Alam M, et al. Isolation of a human anti-HIV gp41 membrane proximal region neutralizing antibody by antigen-specific single B cell sorting. PLoS One, 2011, 6(9): e23532.
  • 3Obiakor H, Sehgal D, Dasso JF, et al. A comparison of hydraulic and laser capture microdissection methods for collection of single B cells, PCR, and sequencing of antibody VDJ. Anal Biochem, 2002, 306(1): 55-62.
  • 4Ozawa T, Kishi H, Muraguchi A. Amplification and analysis of cDNA generated from a single cell by 5'-RACE: application to isolation of antibody heavy and light chain variable gene sequences from single B cells. Biotechniques, 2006, 40(4): 469-470, 472, 474 passim.
  • 5Scheel T, Gursche A, Zacher J, et al. V-region gene analysis of locally defined synovial B and plasma cells reveals selected B cell expansion and accumulation of plasma cell clones in rheumatoid arthritis. Arthritis Rheum, 2011, 63(1): 63-72.
  • 6Menard L, Samuels J, Ng YS, et al. Inflammation-independent defective early B cell tolerance checkpoints in rheumatoid arthritis. Arth Rheum, 2011, 63(5): 1237-1245.
  • 7Jin A, Ozawa T, Tajiri K, et al. A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood. Nat Med, 2009, 15(9): 1088-1092.
  • 8Kurosawa N, Yoshioka M, Isobe M. Target-selective homologous recombination cloning for high-throughput generation of monoclonal antibodies from single plasma cells. BMC Biotechnol, 2011, 11(1): 39.
  • 9Kuppers R, Zhao M, Hansmann ML, et al. Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections. EMBO J, 1993, 12(13): 4955-4967.
  • 10Scheeren FA, van Geelen CM, Yasuda E, et al. Antigen-specific monoclonal antibodies isolated from B cells expressing constitutively active STAT5. PLoS One, 2011, 6(4): e17189.

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