Preparation of Monoclonal Antibody of Malachite Green and Establishment of Direct Competitive ELISA

doi:10.7506/spkx1002-6630-200924061

FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (24 ): 283-286.doi: 10.7506/spkx1002-6630-200924061

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Preparation of Monoclonal Antibody of Malachite Green and Establishment of Direct Competitive ELISA

LI Xiao-li1，CHEN Xue-lan2，LIU Chun-mei3，XIONG Yong-hua1,*

1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China；2. College of Life
Science, Jiangxi Normal University, Nanchang 330022, China ；3. Wuxi Zodolabs Biotech Co. Ltd., Wuxi 214174, China

Received:2009-08-21 Online:2009-12-15 Published:2010-12-29
Contact: XIONG Yong-hua1,* E-mail:yhxiongchen@163.com

Abstract

Abstract:

Carboxyl-malachite green (CMG) was synthesized and immunogen was prepared by coupling CMG with carrier protein BSA through an ester activation method. BALB/c mice were subjected to injection of CMG-BSA and a hybridoma cell line secreting monoclonal antibody against MG was constructed after fusion, screening and cloning experiments. A direct competitive ELISA for detection MG was established with 0.2 μg/ml CMG-HRP and 1:16000 of antibody dilution ratio. This method exhibited high sensitivity and excellent specificity for detecting MG. The standard regression equation of this method was y ＝－20.888x ＋81.423 (R2＝ 0.9813) with a detection limit for MG at the level of 0.05 ng/ml and IC50 was 0.3 ng/ml.

Key words: malachite green, monoclonal antibody, direct competitive ELISA

CLC Number:

LI Xiao-li1，CHEN Xue-lan2，LIU Chun-mei3，XIONG Yong-hua1,*. Preparation of Monoclonal Antibody of Malachite Green and Establishment of Direct Competitive ELISA[J]. FOOD SCIENCE, 2009, 30(24 ): 283-286.

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Preparation of Monoclonal Antibody of Malachite Green and Establishment of Direct Competitive ELISA

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