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1 5 7 Vol. 5 No Journal of Food Safety and Quality Jul., 2014 HTS-ELISA 甄玉萍, 裴世春 *, 高建伟 (, ) 摘要 :,,,,, HTS-ELISA, (Trailing), 关键词 : - ; ; ; Progress of monoclonal antibody secretion hybridoma cell rapid screening technology based on HTS-ELISA ZHEN Yu-Ping, PEI Shi-Chun *, GAO Jian-Wei (College of Food and Biological Engineering, Qiqihar University, Qiqihar , China) ABSTRACT: Various immune analysis technologies based on the monoclonal antibody are the main development direction for modern food safety rapid detection technology. Therefore, preparation of monoclonal antibodies with high-activity has got more and more attention by researchers. Preparation of monoclonal antibody usually involves many techniques, including the preparation of the antigen, immune design, cell fusion, screening of active hybridoma and antibody purification. Among them, the screening of monoclonal antibody secreting hybridoma cells is a key of the prepared monoclonal antibody part. Therefore, in order to provide a reference for high-activity monoclonal antibody preparation and the development of rapid detection technology for food safety, this paper focused on low density fusion cell culture, special equipment and the principle and technology of determining hybridoma cell activity in the screening process according to drag the number of holes (Trailing) to related activities, and mainly introduced one kind of high-activity monoclonal antibody secretion hybridoma technique which was based on HTS-ELISA screening method. KEY WORDS: high-throughput screening ELISA; hybridoma cell; screening; monoclonal antibody 基金项目 : (C201225) ( ) Fund: Supported by the Natural Science Foundation of Heilongjiang Province Science and Technology Department (C201225); and Science and Technology Research Project of Heilongjiang Province Department of Education( ) * 通讯作者 :,, peishichun@qqhru.edu.cn *Corresponding author: PEI Shi-Chun, Professor, College of Food and Biological Engineering, Qiqihar University, No.42, Wenhua Street, Qiqihar , China. peishichun@qqhru.edu.cn

2 7, : HTS-ELISA 引言 [1,2],,, [3] [4,5] [6,7],,,, [8] ELISA (high-throughput screening ELISA, HTS-ELISA) 2 基于 HTS-ELISA 筛选阳性杂交瘤细胞 2.1 常规阳性杂交瘤细胞的筛选, PEG, 96, ELISA [9,10] OD,,,,, OD, ELISA OD, ELISA OD, OD,,, OD, OD,,, [11], ; [12] ; [13],, [14] [15] [16] [17],,,, HTS-ELISA 2.2 HTS-ELISA 筛选方法及特点 (PEG), SP2/0 1:5 1: ,,,,,,, ELISA [18-20] (high-throughput

1962 5 screening ELISA, HTS-ELISA) ELISA 96, HTS-ELISA, SP2/0 1:1, 1 1.5 L 280 μl/ 40 50 96, 96 ( H1 H2 H50), 37 5% CO 2 10 d, 40 50 96,, 280 μl 10 d,,,,, 10 Transtar-96 96 50 μl (C-2, C-2-1 C-2-2.

3 screening ELISA, HTS-ELISA) ELISA 96, HTS-ELISA, SP2/0 1:1, L 280 μl/ , 96 ( H1 H2 H50), 37 5% CO 2 10 d, ,, 280 μl 10 d,,,,, 10 Transtar μl (C-2, C-2-1 C-2-2.C-2-50) (C-1, C-1-1 C-1-2.C-1-50) ELISA ( 1) 1, μl, ELISA, 1, C-1-1 B2 C-2-1 B2, C-1, B2,, ; C-2 E5 D8 H8, C-1,,,, OD, OD (trailing), 2, 96,, 96 (Transtar-96),,,, 2 H8, 2, 3 C-2 H8 OD 2.5,, 4 8 C-2 H8 OD , 3 C-2 H8 4,,,, 4, 3 H8 C-1: 96 + ; H: 96 ; C-2: 96 1 Fig. 1 Positive fusion cell screening model

4 7, : HTS-ELISA 1963 Fig. 2 2 The pattern of Trailing principle, 3 H8 4 H8 HTS-ELISA,, ELISA,,,,, OD,,,,, 3 结束语, HTS-ELISA, 参考文献 [1],,. [J]., 2007, 3(10): Guan H, Shi MJ, Cui YN. Research progress of immunoassay technology[j]. Asia Pacific Tradit Med, 2007, 3(10): [2]. [J]., 2014, 14(3): Wang B. Performance model analysis on detecting markers of HBV by using microparticle enzyme-linked immunosorbent assy[j]. Modern Hospital, 2014, 14(3): [3] Mankotia DS, Irshad M. Cloning and expression of N22 region of Torque Teno virus (TTV) genome and use of peptide in developing immunoassay for TTV antibodies[j]. Virol J, 2014, 11(1): 96. [4],,. [J]., 2013, 42(32): Qi DL, Wang Q, Yi WJ. Preparation identification and application of monoclonal antibodies against histidine-tag[j]. Chongqing Med, 2013, 42(32): [5],,,. [J]., 2011, 30(1): Li K, Sang LY, Bo LJ, et al. Production and Identification of Monoclonal Antibodies to Melamine[J]. J Food Sci Biotechnol, 2011, 30(1): [6],,. [J]., 2010, 26(125): Wang P, Lv XY, Fan SC. Development of the Research and Clinical Application of Genetic Engineering Antibody[J]. J Chongqing Three Gorges Univ, 2010, 26(125): [7],,,. [J].,

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5 , (10): Li J, Lin T, Song S, et al. Advances in Genetic Engineering Antibody[J]. Brotechnol Bull, 2009, (10): [8]. GNA ELISA [D]. :, Shen GE. Preration of Anti-GNA Protein Rabbit Monoclonal Antibodies an Establishment of an Sandwich ELISA basing it[d]. Hangzhou: Zhejiang University, [9],,,. ELISA [J]., 2014, 41(3): Lv N, Yin XP, Zhang HX, et al. Establishment of a Double Antibody Sandwich ELISA for the Detection of Flavobacterium columnaris[j]. China Anim Husbandry Vet Med, 2014, 41(3): [10],,,. M1 ELISA [J]., 2008, 20(5): Pei SC, Zheng LN, Tang YJ, et al. Optimization of ELISA Method Conditions for Detection of AFM1 and it s Application[J]. J Heilongjiang Bayi Agric Univ, 2008, 20(5): [11],,,. :, CN [P] Li PW, Zhang W, Zhang Q, et al. A screening method for hybridoma monoclonal preparation: China, N [P] [12],. :, CN A [P] Gao JE, Wang CX. A method for improving the screening efficiency of hybridoma: China, CN A [P] [13],,,. :, CN A [P] Lu SY, Ren HL, Li YS, et al. A preparation method for marine shellfish toxin monoclonal antibody: China, CN A [P] [14],. :, CN [P] Yan XJ, Guo YH. Method of high-throughput peptide chip for screening of hybridoma cells: China, CN [P] [15]. :, CN [P] Chen GM. A protein chip and its preparation method, application in screening monoclonal antibody: China, CN [P] [16],,. :, CN [P] Zhang QH, Wu C, Miu JM. A polypeptide chip and its preparation method and preparation for monoclonal antibody group: China, CN [P] [17],,. :, CN A [P] Liu Y, Liu J, Zuo W. Preparation of monoclonal antibodies and hybridoma cell strain by various antigens of high throughput: China, CN A [P] [18] Pei SC, Lee WJ, Zhang GP, et al. Development of anti-zearalenone monoclonal antibody and detection of zearalenone in corn products from China by ELISA [J]. Food Contr, 2013, 31: [19],,,. HTS-ELISA M1 [J]., 2010, 50(10): Pei SC, He N, Zhang LJ, et al. Using HTS-ELISA method to prepare anti-aflatoxin M1 monoclonal antibody[j]. Acta Microbiol Sinica, 2010, 50(10): [20],,. [M]. :, Wang SL,Wang YP, Jiang YJ. Food biotechnology[m]. Beijing: Science Press, 作者简介 ( 责任编辑 : 赵静 ) 甄玉萍, 硕士研究生, 主要研究方向为食品营养与安全评估及食品有害物快速检测 @139.com 裴世春, 教授, 主要研究方向为食品营养与安全 peishichun@qqhru.edu.cn

Microsoft Word - 793-797 tb20150504赵宏宇s-高校教改纵横.doc

Microsoft Word - 793-797 tb20150504赵宏宇s-高校教改纵横.doc 微生物学通报 Microbiology China tongbao@im.ac.cn Apr. 20, 2016, 43(4): 793 797 http://journals.im.ac.cn/wswxtbcn DOI: 10.13344/j.microbiol.china.150504 高校教改纵横生物工程专业发酵课程群建设探索 * 赵宏宇