三聚氰胺抗原合成及其多克隆抗体的制备

Synthesis of Melamine Antigen and Preparation of Its Polyclonal Antibodies

以2-氯-4,6-二氨基-1,3,5-三嗪为原料,强碱条件下与3-巯基丙酸反应,制备三聚氰胺半抗原并经质谱和红外鉴定;应用碳二亚胺法将半抗原与BSA和OVA偶联制备三聚氰胺免疫原和包被原,完全抗原经紫外扫描鉴定偶联成功,偶联比分别为16:1和10:1,通过免疫新西兰大白兔成功制备三聚氰胺多克隆抗体,抗体效价可达1:128000以上。本研究为三聚氰胺人工抗原的制备提供了有效途径,为三聚氰胺免疫检测方法的研发奠定了基础。

3-mercaptopropionic acid was reacted with 2-chloro-4, 6-diamino-1, 3, 5-triazine （CAAT） to produce hapten [3- （4, 6-diamino-1, 6-dihydro-1, 3, 5-triazin-2-ylthio） propanoic acid].The molecular structure of the hapetn was identified by mass spectrometry and infrared spectrometry.An immunogen and coating antigen were prepared by coupling the hapten to bovine serum albumin （BSA） or egg ovalbumin （OVA） .The conjugations were identified by UV and animal immunization test. The coupling molar ratios of the hapten and carrier proteins were 16：1 and 10：1 for Mel-BSA and Mel-OVA.Mel -BSA was used to immunize New Zealand white rabbits, and the generation of antibody was detected.The results showed that the titer of antiserum was 1：128 000.The synthesis of MEL artificial antigen was possible to establish a basis for the detection of melamine by immunoassay.