Establishment of Hybridoma Cell Lines Secreting Anti-ractopamine Monoclonal Antibody and Identification of Their Immunological Properties

doi:10.7506/spkx1002-6630-200903039

FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (3): 175-179.doi: 10.7506/spkx1002-6630-200903039

Previous Articles Next Articles

Establishment of Hybridoma Cell Lines Secreting Anti-ractopamine Monoclonal Antibody and Identification of Their Immunological Properties

ZHANG Hai-tang1，WANG Zi-liang1,*，DENG Rui-guang2，ZHONG Hua1，FAN Guo-ying1

(1.College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China；
2.Henan Key Laboratory of Animal Immunology, Zhengzhou 450002, China)

Received:2008-02-22 Revised:2008-05-04 Online:2009-02-01 Published:2010-12-29
Contact: WANG Zi-liang1,* E-mail:wangzl_2008@yahoo.com.cn

Abstract

Abstract:

The active group carboxyl was introduced to ractopamine (RAC) and formed RAC semialdehyde (RAC-SA) which has hapten structure by chemical modification. The method of mixed anhydride (MA) was used to conjugate RAC-SA to bovine serum albumin (BSA) and obtain artificial antigen BSA-RAC identified by infrared (IR), ultraviolet (UV) and SDS-PAGE. Balb/C mice were immunized with BSA-RAC and hybridoma lines that secrete monoclonal antibody against RAC (RAC mAb) were established through cell fusion and then screened using indirect ELISA. Blocking ELISA and the immunological traits of RAC mAb were identified. The results showed that BSA-RAC is synthesized successfully and its conjugation ratio between RAC and BSA is about 24.5:1. Three hybridoma lines of 3F10, 3H12 and 4D8 are screened out and the best one is 4D8 with indirect ELISA titers of 1:1.28×103 in supernatant and 1:6.4×105 in ascites. The isotype of the mAb is IgG1/κ and its affinity constant (Ka) is 1.65×1010L/mol. The mAb shows good sensitivity with an IC50 of 5.7 ng/m lto RAC, 22.3% cross-reactivity with dobutamine and little or no cross-reactivity with other compounds. Establishment of hybridoma lines that secrete RAC mAb makes it possible to develop immunoassay of RAC residues in animal food.

Key words: ractopamine, hapten, artificial antigen, monoclonal antibody, immunological properties

CLC Number:

S859.84

ZHANG Hai-tang1，WANG Zi-liang1,*，DENG Rui-guang2，ZHONG Hua1，FAN Guo-ying1. Establishment of Hybridoma Cell Lines Secreting Anti-ractopamine Monoclonal Antibody and Identification of Their Immunological Properties[J]. FOOD SCIENCE, 2009, 30(3): 175-179.

[1]	WANG Xiaorui, QIU Hongling, WANG Shouli, ZHU Haihua, ZHOU Li, PING Yang, TAN Jing, WANG Yong, WANG Hui. Preparation of Monoclonal Antibodies against Outer Membrane Protein K of Vibrio parahaemolyticus and Development of a Double Antibody Sandwich ELISA [J]. FOOD SCIENCE, 2021, 42(4): 319-325.
[2]	TAN Shu, YANG Jinyi, XU Jihua, SHEN Yudong, ZENG Daoping, SU Xiaona, ZHONG Cuili, XU Xiaoxuan, SUN Yuanming. Development of an Indirect Competitive Enzyme Linked Immunoassay for the Analysis of Amantadine in Chicken Muscle [J]. FOOD SCIENCE, 2020, 41(10): 304-310.
[3]	WANG Liwei, LIU Jin, WU Junrui, WU Rina, YUE Xiqing. Preparation and Identification of Monoclonal Antibody against Bovine Immonoglobulin G [J]. FOOD SCIENCE, 2019, 40(2): 110-116.
[4]	LIU Beibei , CAO Lin. Quantum Dot-Based Assay for Rapid Simultaneous Detection of Lead and Cadmium [J]. FOOD SCIENCE, 2019, 40(18): 335-341.
[5]	XIE Manman, LI Jianwu, WANG Guangbin, ZENG Haijuan, ZHAI Xuzhao, DING Chengchao, LIU Wukang, LIU Qing. Preparation and Immunological Characteristics of Monoclonal Antibodies against Vibrio parahaemolyticus [J]. FOOD SCIENCE, 2017, 38(8): 63-68.
[6]	LI Jie, DING Chengchao, ZHAI Xuzhao, WANG Guangbin, LIU Wukang, XIE Manman, ZENG Haijuan,. Preparation and Characterization of Monoclonal Antibodies against Vibrio Outer Membrane Protein OmpK [J]. FOOD SCIENCE, 2017, 38(6): 111-116.
[7]	WANG Jing, MA Ningning, SONG Yang. Preparation of Artificial Antigen and Polyclonal Antibody for the Detection of Heterocyclic Amines [J]. FOOD SCIENCE, 2017, 38(4): 45-50.
[8]	WANG Jing, WANG Yao, WANG Fangyu, DENG Ruiguang, HU Xiaofei, YU Qiuying, HAO Junfang, XING Yunrui, HOU Yuze. Preparation of Artificial Antigen and ELISA Identification of Rabbit Polyclonal Antiserum for the Detection of Glyphosate [J]. FOOD SCIENCE, 2017, 38(4): 65-69.
[9]	ZHANG Yuanyuan, WANG Ruixin, LI Qiuwei, LI Shuguo. An Immunosensor for Rapid Determination of Ractopamine in Foods Based on Multi-Walled Carbon Nanotubes [J]. FOOD SCIENCE, 2016, 37(8): 170-175.
[10]	WANG Xiaofen, YAN Aiping, TAN Ting, MAO Xuejin, LIU Junjun, GUO Lan, WAN Yiqun. Synthesis and Identification of Stable Artificial Paclobutrazol Antigen [J]. FOOD SCIENCE, 2016, 37(7): 134-139.
[11]	LAI Dan, ZHENG Shengwu, ZHAO Suqing. Synthesis of Artificial Antigens for Immunoassay of Four Phthalate Esters [J]. FOOD SCIENCE, 2016, 37(5): 114-120.
[12]	CHEN Yinnan, CHEN Hua, SHI Xianai, YE Xiaojun, FAN Haiping. Preparation and Application of Monoclonal Antibody against Furazolidone [J]. FOOD SCIENCE, 2016, 37(3): 151-157.
[13]	XIAO Shimei, YAN Aiping, XIAO Fang, WAN Yiqun, GUO Lan. Synthesis and Identification of Artificial Antigens of Forchlorfenuron [J]. FOOD SCIENCE, 2016, 37(21): 183-188.
[14]	WANG Yanan, WANG Xiaofei, NIU Linlin, LEI Zhuang, ZHANG Haitang, WANG Ziliang. Establishment and Preliminary Application of Colloidal Gold Immunochromatography for Detecting Heavy Metal Cadmium Ion in Foods [J]. FOOD SCIENCE, 2016, 37(18): 152-158.
[15]	ZHANG Shuai, QI Yingying, ZHANG Hongxing, WANG Yiwen, XIE Yuanhong, LIU Hui. Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA [J]. FOOD SCIENCE, 2016, 37(16): 211-215.

Establishment of Hybridoma Cell Lines Secreting Anti-ractopamine Monoclonal Antibody and Identification of Their Immunological Properties

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments