抗小鼠PLRP1多克隆抗体的制备以及特异性鉴定和应用

小鼠胰泡细胞表达和分泌一种被称为胰甘油三脂酶(PTL)的脂肪酶，主要参与食物来源的甘油三脂的消化吸收，胰泡细胞同时也表达胰脂肪酶相关蛋白1(PLRP1)，它同PTL具有很高的同源性．为了研究PLRP1的生物学功能，需要制备其抗体．应用谷胱甘肽S-转移酶(GST)表达系统表达了GST融合蛋白，亲和纯化后用以免疫新西兰大白兔，获得了抗PLRP1的多克隆抗体．免疫印迹分析表明，该多克隆抗血清能检测出0.6 ng的融合蛋白抗原以及在3 μg的小鼠胰液提取物中检测出PLRP1蛋白．在证明抗体的特异性方面，尝试了一种新的方法：用PLRP1基因剔除的小鼠作为阴性对照，通过免疫印迹和免疫组化实验证明了该多克隆抗血清具有很高的特异性．进一步的研究发现，进食能促进胰腺中PLRP1的外分泌，这表明PLRP1可能在食物的消化过程中具有一定的生物学功能．

Preparation, Characterization and Application of High Specific Polyclonal Antibody Against Pancreatic Lipase Related Protein 1

Pancreatic acinar cells synthesize pancreatic lipase related protein 1 (PLRPl), which has a high degree of sequence and structural homology with pancreatic triglyceride lipase (PTL). PTL is required for efficient dietary triglyceride digestion, while the physiological role of PLRPl has not been elucidated, although some investigations have shown that its expression level is changed under some physiological or pathological conditions. Specific antigenic peptides were fused to glutathione S-transferase (GST) and purified recombinant fusion protein was used to generate polyclonal antisera by immunization of rabbits. The antisera could detect the antigen as low as 0.6 ng and PLRPI protein in 3 μg of mouse pancreatic juice extracts. The high specificity was verified in Western blot and immunohistochemistry analysis by using PLRPl knockout (KO) mice as the control. Furthermore, it was showed that food intake could increase the exocrine secretion of PLRPl into pancreatic juice. This implied that PLRPl may fulfill dietary digestion function in the digestion track.