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.
Highlights
Fumonisins, which assume significance due to their deleterious effects on animals, are a group of potent, carcinogenic, hepatotoxic and nephrotoxic secondary metabolites produced mainly by Fusarium verticillioides and Fusarium proliferatum [1,2,3].Fumonisin B1 (FB1), the most common and highest toxic of fumonisins species, is the focus of governments and scientists throughout the world due to the strong toxicity and potent carcinogenicity shown in animal studies [4]
The aim of the present study is to find an available method for identifying the conjugation of FB1 to carrier proteins and six commonly used methods are described and compared
GA is utilized as a short cross-linker reagent to aid FB1 linking to bovine serum albumin (BSA) and OVA, respectively
Summary
Fumonisins, which assume significance due to their deleterious effects on animals, are a group of potent, carcinogenic, hepatotoxic and nephrotoxic secondary metabolites produced mainly by Fusarium verticillioides (formerly Fusarium moniliforme Sheldon) and Fusarium proliferatum [1,2,3]. Fumonisin B1 (FB1), the most common and highest toxic of fumonisins species, is the focus of governments and scientists throughout the world due to the strong toxicity and potent carcinogenicity shown in animal studies [4]. FB1 is hepatotoxic and nephrotoxic and it can cause serious diseases in equine species, swine and rodents. Due to its widespread existence in agriculture, the pollution in cereal and products has been reported, which may cause potential health hazards [8,9,10].
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