Preparation and Identification of Monoclonal Antibody for Sarafloxacin

doi:10.7668/hbnxb.2016.02.015

Abstract

Abstract: In order to preparation of sarafloxacin(SARA) monoclonal antibody with good sensitivity, high affinity and strong specificity, synthesized SARA artificial antigen by carbodiimide method.It was used to immunize BALB/c mice.The mice which possessed high serum titer and good sensitivity were selected for cell fusing through cell hybridoma technique.Then to filtrate the hybridoma cell strain which secreted SARA monoclonal antibody(mAb).Prepared SARA mAb by inducing ascites in vivo, and identificated the immunity characteristics of SARA mAb via indirect ELISA and indirect competitive ELISA.The result indicated that after mice were immune by immunogen, their serum antibody titer were all up to 1:128 000.Number 2 mouse with good sensitivity was chosen for cell fusion.After multiple sub cloning, 1G3 and 3H3 hybridoma cell strain were filtered.The titer of SARA mAb from 1G3 was 1:512 000, IC₅₀ was 6.34 ng/mL, affinity constant was 8.55×10⁸ L/mol, cross-reactivity with norfloxacin was 1.06%, cross-reactivity with other drugs were below 0.50%, higher than that of 3H3.

Key words: Sarafloxacin, Monoclonal antibody, Hybridoma cell, Immunological characteristics

CLC Number:

S859.84

LIU Rubiao, HU Xiaofei, ZHANG Gaiping, WANG Hui, WANG Fangyu, LIU Lu, LIU Xi, DENG Ruiguang, SONG Zhaojun. Preparation and Identification of Monoclonal Antibody for Sarafloxacin[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(2): 87-91. doi: 10.7668/hbnxb.2016.02.015.

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http://www.hbnxb.net/EN/Y2016/V31/I2/87

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