Screening and identification of human anti-HIF-1α scFv of lung adenocarcinoma

王洁; 李建国; 庞华; 彭志平; 李少林

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Screening and identification of human anti-HIF-1α scFv of lung adenocarcinoma

更新时间：2026-01-09

- Screening and identification of human anti-HIF-1α scFv of lung adenocarcinoma
- Journal of Army Medical University Vol. 31, Issue 23, Pages: 2319-2322(2009)
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  CLC： R392.11;R392.7;R734.2
- Published：2009
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王洁,李建国,庞华,彭志平,李少林.抗HIF-1α肺腺癌人源单链抗体的筛选和初步鉴定[J].陆军军医大学学报(原第三军医大学学报),2009,31(23):2319-2322.

　WANG Jie,LI Jian-guo,PANG Hua,PENG Zhi-ping,LI Shao-lin.Screening and identification of human anti-HIF-1α scFv of lung adenocarcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(23):2319-2322.
王洁,李建国,庞华,彭志平,李少林.抗HIF-1α肺腺癌人源单链抗体的筛选和初步鉴定[J].陆军军医大学学报(原第三军医大学学报),2009,31(23):2319-2322. DOI：

　WANG Jie,LI Jian-guo,PANG Hua,PENG Zhi-ping,LI Shao-lin.Screening and identification of human anti-HIF-1α scFv of lung adenocarcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(23):2319-2322. DOI：

摘要

目的从单链大容量噬菌体抗体库中筛选特异性的抗HIF-1α肺腺癌人源单链抗体(scFv)，并进行初步鉴定。方法先后以肺腺癌细胞A549、HIF-1α为抗原，通过“吸附-洗脱-扩增”从大容量噬菌体抗体库中筛选特异性抗体，感染大肠杆菌HB2151，实现可溶性抗体的表达，ELISA、免疫细胞化学检测抗原抗体结合活性，SDS-PAGE和Western blot检测可溶性抗体表达和相对分子质量。结果经筛选后，单链抗体得到富集。随机选取10个克隆，ELISA检测5个与HIF-1α抗原呈阳性反应，阳性率为50%。免疫细胞化学表明抗体与A549细胞特异性结合。SDS-PAGE、Western blot证明单链抗体成功可溶表达，且相对分子质量在30×10

左右，ELISA显示其有较好的特异性。结论利用单链大容量噬菌体抗体库技术成功获得特异性抗HIF-1α的肺腺癌人源单链抗体。

Abstract

Objective To screen human anti-hypoxia-inducible factor (HIF)-1α scFv of lung adenocarcinoma from large phage antibody library and identify the positive clones. Methods Panning of large phage antibody library against lung adenocarcinoma cell line A549 and HIF-1α was conducted respectively to select specific antibodies. E. coli HB2151 was infected to induce the expression of soluble scFv. The binding activity and specificity were tested by ELISA and immunocytochemical assay. The expression and relative molecular weight of the soluble scFv was detected by SDS-PAGE and Western blot analysis. Results After panning

the target scFv was enriched

and ELISA resu

lts showed that positive reactions to HIF-1α were detected in 5 of 10 random clones with a positive ratio of 50%. Immunocytochemical analysis showed the specific affinity of the antibodies to A549 cells. The soluble human anti-HIF-1α scFv fragments of lung adenocarcinoma were expressed in E. coli HB2151 and then confirmed by SDS-PAGE. The result of Western blotting showed that the relative molecular weight of the soluble scFv was about 30×10

. The binding activity and specificity were confirmed by ELISA. Conclusion Human anti-HIF-1α scFv of lung adenocarcinoma is successfully obtained with large phage antibody library technique.

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王洁

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李少林

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Related Institution

Department of Epidemiology， Faculty of Military Preventive Medicine， Army Medical University （Third Military Medical University）， Chongqing, PLA Key Laboratory of Respiratory Diseases， Second Affiliated Hospital， Army Medical University （Third Military Medical University）

Institute of Respiratory Diseases, PLA Key Laboratory of Respiratory Diseases， Second Affiliated Hospital， Army Medical University （Third Military Medical University）

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