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Int. J. Mol. Sci. 2012, 13(1), 84-96; doi:10.3390/ijms13010084

Characterization and Comparison of Fumonisin B1-Protein Conjugates by Six Methods

College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China
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Received: 10 October 2011 / Revised: 8 November 2011 / Accepted: 28 November 2011 / Published: 22 December 2011
(This article belongs to the Special Issue Protein Conjugates and Bioconjugate Chemistry)
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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 Fumonisin B1; conjugated antigen; carrier protein; molar ratio; antibody
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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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.

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