Characterization of fumonisin B_1 artificial antigen and production of the antibody

In the study,the hapten fumonisin B1(FB1)was coupled with carrier proteins ovalbumin(OVA)and bovine serum albumin(BSA)respectively,by one step method with glutaraldehyde(GA)method,and two artificial antigens FB1-OVA and FB1-BSA were synthesized.Four methods of gel electrophoresis,ultraviolet spectroscopy(UV),infrared spectroscopy(IR)and matrix-assisted laser absorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS)were used to determine the coupled effects and coupling ratios of the artificial antigens.Then FB1-BSA was used to immunize BALB/c mice,and FB1-OVA was used as solid phase coating antigen.The indirect enzyme-linked immunosorbent assay(i-ELISA)was used to determine the titer of anti-serum from mice.The results showed that four methods all proved that carrier proteins coupled with FB1 successfully.The molecular weights of FB1-OVA and FB1-BSA were about 50 000 and 75 000 respectively by gel electrophoresis.The accurate molecular weights of FB1-OVA and FB1-BSA were 48 009.212 and 74 355.301 and the coupling ratios of FB1-OVA and FB1-BSA were 5∶1 and 10∶1 respectively by MALDI-TOF-MS.The titer of anti-serum was 1∶1.25×105 by i-ELISA with good sensitivity.