兔抗人肥大细胞羧肽酶抗体的制备与鉴定

目的：以原核表达的人肥大细胞羧肽酶（hMC-CP）免疫家兔，制备兔抗hMC-CP多克隆抗体并进行鉴定。方法：在大肠杆菌中诱导表达重组hMC—CP，经Ni-NTA—Agarose柱层析纯化后，以其为免疫原制备兔抗hMC-CP多抗，并以间接ELISA测定抗体效价，以Western blot鉴定抗体的特异性。结果：成功地表达并纯化了hMC-CP，以纯化的重hMC-CP组免疫家兔成功制备了兔抗hMC-CP抗体，ELISA结果显示效价达1：6400，Western blot分析表明该抗体能特异结合hMC—CP。结论：以纯化的重组hMC—CP为免疫原，成功地制备了效价、特异性较强兔抗hMC-CP抗体，为进一步建立简便的hMC-CP检测方法及进一步研究hMC-CP生物学功能打下了良好的基础。

Preparation and characterization of rabbit antibody against human mast cell carboxypeptidase

AIM: To prepare antibody against human mast cell carboxypeptidase (hMC-CP) by using recombinant hMC-CP expressed in E.coli, and to characterize the antibody. METHODS: hMC-CP was expressed in E.coli with L-Arabinose induction and purified through Ni-NTA column. The purified hMC-CP as immunogen was used to immunize rabbit. The titer and the specificity of the rabbit anti-hMC-CP antibody was analyzed by indirect ELISA and Western blot respectively. RESULTS: The hMC-CP was successfully expressed and purified. The polyconal anit-hMC-CP antibody was prepared by immunizing rabbit using the purified recombinant protein, The titer of the generated antiserum was 1:6 400 by ELISA. Western blot analysis showed that this antibody could bind with hMC-CP specifically. CONCLUSION: The rabbit anti-hMC-CP antibody with high titer and high specificity has been prepared by using purified recombinant hMC-CP as immunogen, which lays the foundation for further research on detection and function of hMC-CP.