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Characterization and Comparison of Fumonisin B1-Protein Conjugates by Six Methods
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China
* Author to whom correspondence should be addressed.
Received: 10 October 2011; in revised form: 8 November 2011 / Accepted: 28 November 2011 / Published: 22 December 2011
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 B1, FB1) 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 (FB1-OVA and FB1-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 FB1-BSA and FB1-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 FB1-BSA and FB1-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.
Keywords: Fumonisin B1; conjugated antigen; carrier protein; molar ratio; antibody
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MDPI and ACS Style
Wang, Y.; He, C.-H.; Zheng, H.; Zhang, H.-B. Characterization and Comparison of Fumonisin B1-Protein Conjugates by Six Methods. Int. J. Mol. Sci. 2012, 13, 84-96.
Wang Y, He C-H, Zheng H, Zhang H-B. Characterization and Comparison of Fumonisin B1-Protein Conjugates by Six Methods. International Journal of Molecular Sciences. 2012; 13(1):84-96.
Wang, Ying; He, Cheng-Hua; Zheng, Hao; Zhang, Hai-Bin. 2012. "Characterization and Comparison of Fumonisin B1-Protein Conjugates by Six Methods." Int. J. Mol. Sci. 13, no. 1: 84-96.