Construction and characterization of an anti-prion scfv fusion protein pair for detection of prion protein on antibody chip
文献类型：期刊论文
| 作者 | Zhang, J. -B.; Zhang, X. -E.; Zhou, Y. -F.; Bi, L. -J.; Zhang, Z. -P.; Wang, S. -H.; Chen, Y. -Y.; Guo, Y. -C.; Wen, J. -K.; Yu, Z. -N. |
| 刊名 | Analytical letters
 |
| 出版日期 | 2007 |
| 卷号 | 40期号:5页码:855-873 |
| 关键词 | Bovine prion protein Scfv Phage display Fusion protein Antibody chip |
| ISSN号 | 0003-2719 |
| DOI | 10.1080/00032710701242048 |
| 通讯作者 | Zhang, x. -e.(zhangxe@most.cn) |
| 英文摘要 | A pair of single chain fv fragment (scfv) fusion proteins were constructed and characterized. antibody chips using the pair were designed for sensitive detection of prion protein. phage displayed antibody library was synthesized by immunizing mice with thioredoxin-mature bovine prion fusion protein (trxa-bprp(c)). after five rounds of panning against recombinant bovine prion protein (rb-prpc) and elisa test, two positive clones with high affinity to rb-prpc, named z163 and z186, were obtained. they were conjugated with a linker-streptavidin binding protein (sbp) or human igg1 constant fragment (fc) to form the scfv fusion protein pair z186-l-sbp/z163-fc. western blot experiments showed that the scfv fusion pair specifically interacted with the line epitopes of the protease resistant core region bprp27-30. surface plasmon resonance (spr) sensorgrams revealed that the equilibrium dissociation constants of the interactions with rb-prpc were 3.24 x 10(-8) m, 8.82 x 10(-8) m, and 8.10 x 10(-9) m for z186-l-sbp, z163, and z163-fc, respectively. all binding reactions followed rapid association and slow dissociation kinetics. as a detection pair, z186-l-sbp functioned as a capture probe and was immobilized on the streptavidin coated slides to form reactive layer of the antibody chip, and z163-fc labeled with fluorescence dye cy3 functioned as a detection probe generating fluorescence signal. the antibody chip could detect existence of rb-prpc with detection limit of 1 pg/ml. |
| WOS关键词 | MONOCLONAL-ANTIBODIES ; IMMUNOASSAY ; PEPTIDE ; PHAGE ; PRPSC ; IMMOBILIZATION ; MICROARRAYS ; DIAGNOSIS ; SCRAPIE ; LIBRARY |
| WOS研究方向 | Chemistry |
| WOS类目 | Chemistry, Analytical |
| 语种 | 英语 |
| WOS记录号 | WOS:000245466800002 |
| 出版者 | TAYLOR & FRANCIS INC |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2375393 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Zhang, X. -E. |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China 2.Huazhong Agr Univ, Coll Life Sci & Technol, Wuhan, Peoples R China 3.Ctr Dis Prevent & Control Hubei Prov, Wuhan, Peoples R China 4.Chinese Acad Sci, Inst Biophys, State Key Lab Biomacromol, Beijing 100080, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, J. -B.,Zhang, X. -E.,Zhou, Y. -F.,et al. Construction and characterization of an anti-prion scfv fusion protein pair for detection of prion protein on antibody chip[J]. Analytical letters,2007,40(5):855-873. |
| APA | Zhang, J. -B..,Zhang, X. -E..,Zhou, Y. -F..,Bi, L. -J..,Zhang, Z. -P..,...&Yu, Z. -N..(2007).Construction and characterization of an anti-prion scfv fusion protein pair for detection of prion protein on antibody chip.Analytical letters,40(5),855-873. |
| MLA | Zhang, J. -B.,et al."Construction and characterization of an anti-prion scfv fusion protein pair for detection of prion protein on antibody chip".Analytical letters 40.5(2007):855-873. |
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来源：武汉病毒研究所
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