Wang Wei, Yin Xiaotao, Li Yunqi, Tian Renli, Wu Hao, Yan Jinqi, Gao Jiangping, Yu Jiyun
Department of Urology, General Hospital of PLA, Beijing 100853, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 May;29(5):481-4.
To obtain β-chain human chorionic gonadotropin (β-hCG) fusion protein (β-hCG/GST) and identify its antigenicity.
The full-length gene of β-hCG was amplified by PCR. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-β-hCG, and then it was transformed into BL21 (DE3) for β-hCG/GST fusion protein expression under IPTG induction. After SDS-PAGE assay, the fusion protein was purified by affinity chromatography and identified by Western blotting. The antigenicity of the purified fusion protein was characterized by ELISA.
The β-hCG gene we obtained had an identical sequence to that retrieved in GenBank. The prokaryotic expression vector pET-42a-β-hCG was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated that the purified β-hCG fusion protein had satisfactory antigenicity.
The purified β-hCG/GST fusion protein with satisfactory antigenicity has been obtained, which will facilitate further study on active anti-tumor immunotherapy targeting β-hCG.
获取β链人绒毛膜促性腺激素（β-hCG）融合蛋白（β-hCG/GST）并鉴定其抗原性。
通过PCR扩增β-hCG全长基因。将PCR产物克隆至pET-42a原核表达载体构建重组质粒pET-42a-β-hCG，然后转化至BL21（DE3）中，在IPTG诱导下表达β-hCG/GST融合蛋白。经SDS-PAGE分析后，通过亲和层析纯化融合蛋白，并进行Western印迹鉴定。采用ELISA法鉴定纯化融合蛋白的抗原性。
我们获得的β-hCG基因序列与GenBank中检索到的序列一致。经酶切和DNA测序证实，成功构建了原核表达载体pET-42a-β-hCG。Western印迹和ELISA均表明，纯化的β-hCG融合蛋白具有良好的抗原性。
已获得具有良好抗原性的纯化β-hCG/GST融合蛋白，这将有助于进一步开展针对β-hCG的活性抗肿瘤免疫治疗研究。
