Synthesis of Artificial Antigen and Preparation of Polyclonal Antibody for Malachite Green

doi:10.7506/spkx1002-6630-201107061

FOOD SCIENCE ›› 2011, Vol. 32 ›› Issue (7 ): 282-285.doi: 10.7506/spkx1002-6630-201107061

• Bioengineering • Previous Articles Next Articles

Synthesis of Artificial Antigen and Preparation of Polyclonal Antibody for Malachite Green

WANG Mei1，XU Nai-feng2，LIU Li-qiang2，KUANG Hua2，LIN Fei2，SONG Shan-shan2，WANG Ze-lu2，XU Chuan-lai2,*

1. Analytical and Testing Center, Suzhou University, Suzhou 215123, China；
2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

Received:2010-07-16 Revised:2011-03-01 Online:2011-04-15 Published:2011-03-30

Abstract

Abstract: In order to determine malachite green (MG) residues in food, the synthesis of antigen and preparation of polyclonal antibody for MG were explored. In this study, carboxylic malachite green (CMG) hapten was first synthesized and analyzed by LC-MS. The hapten was coupled separately with the two carrier proteins BSA and OVA by diazotization method, and identified by UV-visible spectroscopy method. Compared with CMG standard, BSA and OVA, the absorption peaks CMG-BSA and CMG-OVA displayed a marked shift, indicating that the successful conjugation between CMG and BSA or OVA, and the coupling ratio of the immunogen CMG-BSA was calculated to be 10.6:1. Polyclonal antibody for MG was prepared by immunization with CMG-BSA and evaluated using the titer of anti-CMG antisera determined by indirect ELISA. Furthermore, two rabbits were used to evaluate antibody specificity with determined IC50 values of 8.1 ng/mL and 4.6 ng/mL, respectively.

Key words: malachite green (MG), antigen, polyclonal antibody, indirect ELISA

CLC Number:

R392.11

WANG Mei1，XU Nai-feng2，LIU Li-qiang2，KUANG Hua2，LIN Fei2，SONG Shan-shan2，WANG Ze-lu2，XU Chuan-lai2,*. Synthesis of Artificial Antigen and Preparation of Polyclonal Antibody for Malachite Green[J]. FOOD SCIENCE, 2011, 32(7 ): 282-285.

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Synthesis of Artificial Antigen and Preparation of Polyclonal Antibody for Malachite Green

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