Characterization and Comparison of Ochratoxin A-Ovalbumin （OTA-OVA） Conjugation by Three Methods
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-fl ight 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 effi cient methods to evaluate the cross-linking effect and calculate the molecular coupling ratio.