Characterization and comparison of Fumonisin B(1)-protein conjugates by six methods.
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.
College 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.).
Ying
Y
Wang
Cheng-Hua
CH
He
Hao
H
Zheng
Hai-Bin
HB
Zhang
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D037341:Fumonisins / Q000737:chemistry*; D005976:Glutaral / Q000737:chemistry; D006241:Haptens / Q000737:chemistry; D008970:Molecular Weight; D010047:Ovalbumin / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017550:Spectroscopy, Fourier Transform Infrared
D037341:Fumonisins; D006241:Haptens; D012710:Serum Albumin, Bovine; C056933:fumonisin B1; D010047:Ovalbumin; D005976:Glutaral