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Characterization and comparison of Fumonisin B(1)-protein conjugates by six methods
Ying Wang1, Cheng-Hua He, Hao Zheng
1College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China; E-Mails: wangying0624@hotmail.com (Y.W.); hechenghua@njau.edu.cn (C.-H.H.); zebeihao@yahoo.com.cn (H.Z.).
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 (Fumonisin B(1), FB(1)) was coupled to ovalbumin (OVA) and bovine serum albumin (BSA), respectively by a short cross-linker reagent (glutaraldehyde, GA). To develop a technique for detecting the conjugation, the hapten-protein conjugates (FB(1)-OVA and FB(1)-BSA) were characterized thoroughly by ultraviolet (UV) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. The molecular weights of FB(1)-BSA and FB(1)-OVA were 74,355.301 Da and 48,009.212 Da, respectively determined by the method of MALDI-TOF-MS. The molecular coupling ratios were 11 and 5 in FB(1)-BSA and FB(1)-OVA, respectively. In this experiment, MALDI-TOF-MS was selected as the most efficient method to evaluate the cross-linking effect and calculate the molecular coupling ratio.
