切胶免疫制备腮腺液高丰度蛋白多克隆抗体

目的 通过切胶免疫制备人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备提供抗原解决方案.方法 将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量约为50-65kDa的高丰度蛋白条带,研磨后注射新西兰大耳白兔诱导产生免疫应答并制备兔抗人腮腺液高丰度蛋白多克隆抗体,并应用蛋白免疫印记(Western blot)等方法进行抗体鉴定.结果 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体.结论 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备及唾液蛋白质组学研究奠定基础.

Preparation and characterization of polyclonal antibodies against high abundant proteins in human parotid saliva

Objective To prepare and characterize polyclonal antibodies against high abundant proteins in human parotid saliva for the preparation of monoclonal antibodies(mAbs).Methods Proteins in human parotid saliva were concentrated by using ultrafiltration and analyzed by SDS-PAGE.The proteins strap between 50-65kDa was cut,grinded and used to immunize New Zealand rabbits.The polyclonal antibodies against high abundant proteins in human parotid saliva were prepared and characterized by Western blot.Results Polyclonal antibodies against high abundant proteins in human parotid saliva were successfully prepared and characterized.Conclusion Polyclonal antibodies against high abundant proteins in human parotid saliva were successfully prepared and characterized.The present study may provide a premise in the preparation of mAbs against high abundant proteins and future study of human saliva proteome.