目的:合成玉米赤霉烯酮(ZEN)半抗原及全抗原,并鉴定其是否合成成功,为制备ZEN抗体奠定基础。方法:将ZEN与羧甲氧基胺半盐酸盐(O-(carboxym ethyl)hydroxyla-m ine hem ihydrochloride)反应,合成半抗原ZEN-6-羧甲氧基胺(ZEN-oxim e),采用...目的:合成玉米赤霉烯酮(ZEN)半抗原及全抗原,并鉴定其是否合成成功,为制备ZEN抗体奠定基础。方法:将ZEN与羧甲氧基胺半盐酸盐(O-(carboxym ethyl)hydroxyla-m ine hem ihydrochloride)反应,合成半抗原ZEN-6-羧甲氧基胺(ZEN-oxim e),采用TLC、HPLC和液相-质谱联用技术进行半抗原的纯化、鉴定;通过活泼酯法将半抗原与载体蛋白偶联制备ZEN全抗原,采用紫外光谱法、TNBS法结合比的测定以及免疫学方法进行全抗原鉴定。结果:鉴定结果表明目标半抗原及全抗原合成成功。结论:本研究采用多种方法进行半抗原及全抗原的鉴定,结果皆表明目标半抗原及全抗原合成成功,说明本研究合成ZEN半抗原及人工抗原的技术路线是可行的,为进一步研制ZEN的抗体奠定良好基础。展开更多
The thiol group of glutathione (GSH) was protected by 2, 4-dinitrochlorobenzene(DNCB). The product GSH-S-DNP was reacted with 2-methyl-1-propanol and 3-methyl-1-butanol respectively to obtain hapten6 and 7. Their stru...The thiol group of glutathione (GSH) was protected by 2, 4-dinitrochlorobenzene(DNCB). The product GSH-S-DNP was reacted with 2-methyl-1-propanol and 3-methyl-1-butanol respectively to obtain hapten6 and 7. Their structures were determined by 400 MHz1H NMR and IR. The two haptens were then covalently crosslinked with bovine serum albu-min(BSA) hy using glutaraldehyde as coupling agent to obtain antigen 6 and 7. The resultsof gel electrophoresis analysis gave the average molecular weight of 1. 27 ×10-22 kg for eachantigen. The average connected ratio of BSA to hapten was 1: 12. Other UV and fluores-cence spectral properties of the antigens are reported.展开更多
Three thyroxine haptens were synthesized by protecting amino group of thyroxine with Cu 2+ ion and characterized by 1H NMR, MS, elemental analysis and IR. Thyroxine and the derivatives were coupled with carrier protei...Three thyroxine haptens were synthesized by protecting amino group of thyroxine with Cu 2+ ion and characterized by 1H NMR, MS, elemental analysis and IR. Thyroxine and the derivatives were coupled with carrier proteins(BSA and Ovalbumin). Thus antigens were prepared and hapten quantities of the antigens were measured by UV spectroscopy at 320 nm. An anti O methyl T 4 monoclonal antibody was prepared by immunization, cell fusion and screening.展开更多
文摘目的:合成玉米赤霉烯酮(ZEN)半抗原及全抗原,并鉴定其是否合成成功,为制备ZEN抗体奠定基础。方法:将ZEN与羧甲氧基胺半盐酸盐(O-(carboxym ethyl)hydroxyla-m ine hem ihydrochloride)反应,合成半抗原ZEN-6-羧甲氧基胺(ZEN-oxim e),采用TLC、HPLC和液相-质谱联用技术进行半抗原的纯化、鉴定;通过活泼酯法将半抗原与载体蛋白偶联制备ZEN全抗原,采用紫外光谱法、TNBS法结合比的测定以及免疫学方法进行全抗原鉴定。结果:鉴定结果表明目标半抗原及全抗原合成成功。结论:本研究采用多种方法进行半抗原及全抗原的鉴定,结果皆表明目标半抗原及全抗原合成成功,说明本研究合成ZEN半抗原及人工抗原的技术路线是可行的,为进一步研制ZEN的抗体奠定良好基础。
文摘The thiol group of glutathione (GSH) was protected by 2, 4-dinitrochlorobenzene(DNCB). The product GSH-S-DNP was reacted with 2-methyl-1-propanol and 3-methyl-1-butanol respectively to obtain hapten6 and 7. Their structures were determined by 400 MHz1H NMR and IR. The two haptens were then covalently crosslinked with bovine serum albu-min(BSA) hy using glutaraldehyde as coupling agent to obtain antigen 6 and 7. The resultsof gel electrophoresis analysis gave the average molecular weight of 1. 27 ×10-22 kg for eachantigen. The average connected ratio of BSA to hapten was 1: 12. Other UV and fluores-cence spectral properties of the antigens are reported.
文摘Three thyroxine haptens were synthesized by protecting amino group of thyroxine with Cu 2+ ion and characterized by 1H NMR, MS, elemental analysis and IR. Thyroxine and the derivatives were coupled with carrier proteins(BSA and Ovalbumin). Thus antigens were prepared and hapten quantities of the antigens were measured by UV spectroscopy at 320 nm. An anti O methyl T 4 monoclonal antibody was prepared by immunization, cell fusion and screening.
