金免疫层析法检测盐酸克伦特罗的工艺研究-生物化工专业论文.docxVIP

ABSTRACT W批h the development of immunization technique and the emerging food security issues，gold immunochromatography assay for its particular competitive advantages is playing an increasingly important role in the clinical，field testing，也mily selιtest， and so on. However，there exists a considerable technical gap between domestic and foreign. This article aims to establish R D technology platform of it from antigen h呗design to sensitivity adjusting through the technological study of clenbuterol h呗 ··p匈The frrst step is antigen design according ωthe molecular characteristics of clenbuterol. Complete antigen used in the current commercial products was mostly synthesized by diazoti泣z剖atio忏n1-cωoupl怕ingme创thοd耐(OVA ··p 匈 But 批 i阳ss町y削nlt协hes阳泣e叫d by cross-linking with gl川u此ta即r剖al岱deh巧ψyyde in this a眈rt削icl峭e叫(OVA-GA-CL， coupling ratio of 纣25.止2:1). Immune response was more easily tlourished ，because the fivecarbon bridg reduces the coupling protein steric effect. Meanwhile ，polyclonal antibody w:础 purified by affmity chromatography and ammonium sulfate precipitation in this article and the antibody titer was detected by indirect ELISA.ηle purified antibody titers were respectively 1:16000 and 1:32000. Compared to the antiserum， the purified antibody titer was decreased ，the analysis result of SDS showed that the purity of purified antibody raised greatl弘 and the strips were prepared on this basis. Secondl)与 colloidal gold solution was prepared by improved Frens method. Reaction system was heated by water bath with rotary evaporato民The sodium citrate was fistly added to the systerm and after that the chlorine acid was mixed in the odium citrate solution. In the end，single scattered small particles were prepared and ‘蝇 were identified by transmission electron microscopy and full wavelength scann侃 u At last，through the optimization of the pretreatment solution of the sample pad ， and