Ren Qian, Liu Yong-Jun, Xu Bin, Han Zhi-Bo, Lu Shi-Hong, Ma Feng-Xia, Chen Zhong, Han Zhong-Chao
State Key Laboratory of Experimental Hematology, National Research Center for Stem Cell Engineering and Technology, CAMS and PUMC, Tianjin 300020, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Nov;22(6):801-3.
To express the recombinant fusion protein of hemangiopoietin (HAPO) and prepare the rabbit-anti-human HAPO polyclonal antibody.
The sequence encoding HAPO was amplified by PCR and cloned into plasmid pET32c to construct recombinant prokaryotic expression system. The recombinant expression vectors were identified by enzyme digestion analysis and transformed into E. coli. The HAPO protein was purified by affinity chromatography. Rabbits were immunized with the HAPO protein, and the immune sera of rabbits were collected. Antibodies (IgG) obtained from the immune sera were purified.
The purified HAPO protein was successfully obtained. The purified polyclonal antibody of rabbit-anti-human HAPO was also obtained from the immune sera of rabbits, and could response to human HAPO.
A prokaryotic expression system of human HAPO has been prepared and the polyclonal antibody against HAPO has been prepared, which can be used to determine HAPO protein.
表达血管生成素（HAPO）重组融合蛋白并制备兔抗人HAPO多克隆抗体。
通过PCR扩增编码HAPO的序列，并克隆到质粒pET32c中构建重组原核表达系统。通过酶切分析鉴定重组表达载体，并转化到大肠杆菌中。通过亲和层析纯化HAPO蛋白。用HAPO蛋白免疫兔子，收集兔子的免疫血清。从免疫血清中获得的抗体（IgG）进行纯化。
成功获得纯化的HAPO蛋白。从兔子免疫血清中也获得了纯化的兔抗人HAPO多克隆抗体，且能与人HAPO发生反应。
已制备出人HAPO的原核表达系统，并制备了抗HAPO多克隆抗体，可用于检测HAPO蛋白。
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