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| Characterization and Comparison of Ochratoxin A-Ovalbumin (OTA-OVA) Conjugation by Three Methods |
| WANG Xi-chun, BAO Ming, WU Jin-jie, LUO Ying, MA Liang-you, WANG Ying, ZHANG Ai-hua, HE Cheng-hua , ZHANG Hai-bin |
1、College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, P.R.China
2、Aquatic Technology Promotion Station of Anhui Province, Hefei 230066, P.R.China
3、College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.China |
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摘要 In order to generate an antibody against a small hapten molecule, the hapten is cross-linked with carrier protein to make it immunogenic. In this study, the hapten (ochratoxin A, OTA) was coupled to ovalbumin (OVA) by an active ester reaction. To develop a technique for detecting the conjugation, the hapten-protein conjugate (OTA-OVA) was characterized thoroughly by immunoarray technology, ultraviolet (UV) spectroscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. The molecular weight of OTA-OVA was 50 350.141 Da, and the molecular weight of OVA was 44 887.506 Da, which were determined by MALDI-TOF-MS, respectively. In OTA-OVA, the molecular coupling ratio was 13:1 by MALDI-TOF-MS while the molecular coupling ratio was 10:1 by UV. In this experiment, UV and MALDI-TOF-MS were selected as the efficient methods to evaluate the cross-linking effect and calculate the molecular coupling ratio.
Abstract In order to generate an antibody against a small hapten molecule, the hapten is cross-linked with carrier protein to make it immunogenic. In this study, the hapten (ochratoxin A, OTA) was coupled to ovalbumin (OVA) by an active ester reaction. To develop a technique for detecting the conjugation, the hapten-protein conjugate (OTA-OVA) was characterized thoroughly by immunoarray technology, ultraviolet (UV) spectroscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. The molecular weight of OTA-OVA was 50 350.141 Da, and the molecular weight of OVA was 44 887.506 Da, which were determined by MALDI-TOF-MS, respectively. In OTA-OVA, the molecular coupling ratio was 13:1 by MALDI-TOF-MS while the molecular coupling ratio was 10:1 by UV. In this experiment, UV and MALDI-TOF-MS were selected as the efficient methods to evaluate the cross-linking effect and calculate the molecular coupling ratio.
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Received: 01 November 2012
Accepted:
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| Fund: Acknowledgements This work was supported by the National High-Technology Research and Development Program of China (2007AA10Z429) and the Stabling and Introducing Talents Fund Program of Anhui Agricultural University, China (WD2011-17). |
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Corresponding Authors:
WU Jin-jie, Tel: +86-551-5786329, Fax: +86-25-5785027, E-mail: wjj@ahau.edu.cn
E-mail: wjj@ahau.edu.cn
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| About author: WANG Xi-chun, E-mail: wangxichun@ahau.edu.cn; |
Cite this article:
WANG Xi-chun, BAO Ming, WU Jin-jie, LUO Ying, MA Liang-you, WANG Ying, ZHANG Ai-hua, HE Cheng-hua , ZHANG Hai-bin.
2014.
Characterization and Comparison of Ochratoxin A-Ovalbumin (OTA-OVA) Conjugation by Three Methods. Journal of Integrative Agriculture, 13(5): 1130-1136.
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