人类ARL-1重组蛋白特异性抗体的制备及鉴定

The Preparation and Identification of Human ARL-1 Recombinant Protein and Its Specific Antibody

目的 制备ARL -1重组蛋白及其特异性抗体。方法 利用基因重组技术构建PQE -ARL -1原核表达载体 ,在M15大肠杆菌中进行表达 ,制备ARL -1重组蛋白 ,再将此蛋白免疫健康新西兰兔 ,制备ARL -1特异性抗体。结果 ARL -1在大肠杆菌中获得高效表达 ,经Ni+-NTA (次氮基三乙酸 )琼脂柱纯化 ,获得纯蛋白 ,将此蛋白免疫健康新西兰兔 ,制备出抗人ARL-1特异性抗体 ,经环状沉淀试验及双琼脂扩散试验检测出抗体效价达 1∶2 0 0 0 0。结论 人类ARL -1重组蛋白及其特异性抗体的制备 ,为进一步研究ARL -1在肝癌早期诊断中的作用 。

Objective To prepare the ARL 1 recombinant protein and its specific antibody.Methods The ARL 1 cDNA was cloned into procaryotic expression vector PQE 30 and the recombinant ARL 1 was expressed in E.coli M15,The recombinant protein of ARL 1 was prepared;This protein was then used to immunize the healthy New Zealand rabbits to obtain the specific antibody against ARL 1.Results The recombinant protein of ARL 1 was expressed effectively in E.coli.After purified with Ni + NTA,the protein was used to immunize the healthy New Zealand rabbits to prepare the specific antibody against ARL 1.The titer of the antibody was 1∶20 000 according to the ring precipitation test and the double agar diffusion precipitation test.Conclusion The preparation of human ARL 1 recombinant protein and its specific antibody facilitates a possibility for further study of ARL 1 in the early diagnosis of liver cancer.This antibody can also be used to carry nadiate isotopas or drugs that resist tumour for taget killing of the liver cancer cells.