Characterization and Comparison of Ochratoxin A-Ovalbumin （OTA-OVA） Conjugation by Three Methods

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, th...

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Published in:农业科学学报：英文版 2014 (5), p.1130-1136
Main Author: WANG Xi-chun BAO Ming WU Jin-jie LUO Ying MA Liang-you WANG Wing ZHANG Ai-hua HE Cheng-hua ZHANG Hai-bin
Format: Article
Language:English
Subjects:
MALDI-TOF-MS
共轭
半抗原
卵清蛋白
基质辅助激光解吸电离质谱
抗原分子
表征
赭曲霉毒素A
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Summary: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.
ISSN:2095-3119
2352-3425