5 7 Vol. 5 No. 7 2014 7 Journal of Food Safety and Quality Jul., 2014 HTS-ELISA 甄玉萍, 裴世春 *, 高建伟 (, 161006) 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 161006, 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) (12541871) 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(12541871) * 通讯作者 :,, E-mail: peishichun@qqhru.edu.cn *Corresponding author: PEI Shi-Chun, Professor, College of Food and Biological Engineering, Qiqihar University, No.42, Wenhua Street, Qiqihar 161006, China. E-mail: peishichun@qqhru.edu.cn
7, : HTS-ELISA 1961 1 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 1:10 5-10 96,,,,,,, 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.C-2-50) (C-1, C-1-1 C-1-2.C-1-50) ELISA ( 1) 1, 10 96 50 μl, ELISA, 1, C-1-1 B2 C-2-1 B2, C-1, B2,, 40 50 ; 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 2.3 1.9 1.1 0.5, 3 C-2 H8 4,,,, 4, 3 H8 C-1: 96 + ; H: 96 ; C-2: 96 1 Fig. 1 Positive fusion cell screening model
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