氯霉素全抗原的合成及其卵黄抗体的分离纯化-生物化工专业毕业论文.docxVIP

氯霉素全抗原的合成及其卵黄抗体的分离纯化Abstract 氯霉素全抗原的合成及其卵黄抗体的分离纯化 Abstract Chloramphenicol is all antibiotic with a broad spectrum．It is danger to human health due to its toxicity,so the residual chloramphenicol must be stirictly controlled and detected in food and animal production Enzyme—linked immunosorbent assay(ELISA)methods offer the possibility of highly sensitive，relatively rapid and cost—effective measurements． ELISAs are extensively studied and applied in pesticide residue and toxin monitoring， which is an area with enormous potential for growth． In order to establish an enzyme-linked immunosorbent assay(ELISA)for mpidly examing chloramphenicol(CAP)，the antibody衍tIl lligh titer and specificity must be firstly prepared．So it is necessary for antibody production to synthesize an immunogen by conjugating the hapten(CAP)with a carrier protein．CAP was coupled with bovine serum albumin(BSA)to prepare a complete antigen CAP-BSA by glutaraldehyde(GA) and N—N’一carbonyldiimidazole(CDI)in this paper．After dialysis or ultrafiltration，the products were purified with Superdex 75．The target product was obtained by collecting the right first fraction in the wash curve from the gel filtration．The successful linkage of the CAP—BSA was identified by the ultraviolet spectrum(uv)and the infrared spectrometry(IRS)．The residual amount of amino groups was determined by the TNBS spectrometry,and the amount of CAP linked to BSA was achieved．A large amount of CAP was coupled in the complete antigen by glutaraldehyde method，while an appropriate amountby N—N’一carbonyldiimidazole(CDI)method，mostly between ten and twenty．SDS．polyacrylamide gel electrophoresis was used to analyse the purified products．There existed only one zone in SDS—PAGE and the products can be considered as an immunogen．The content of CAP in the CAP-BSA had been determined by an ELISA kit purchased from market． All the above results indicated that the immunogen CAP—BSA was successfully synthesized，which was going to be injected in t