A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China | |
Zhang, JY; Wang, JG; Wu, YS; Li, M; Li, AH; Gong, XL | |
刊名 | JOURNAL OF FISH DISEASES
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2006 | |
卷号 | 29期号:1页码:1-7 |
关键词 | Carassius auratus China combined antibody library Myxobolus rotundus phage display single-chain antibody fragments |
ISSN号 | 0140-7775 |
通讯作者 | Wang, JG, Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China |
中文摘要 | Immunological methods have been developed for the diagnosis of Myxobolus rotundus but their use has been limited for the prevention and therapy of this serious parasitic pathogen. Phage display antibody libraries are a powerful technique for the development of antibodies to molecules of interest and have advantages over traditional hybridroma approaches. In the present study, four antigen fractions related to M. rotundus were prepared and a combined phage display single-chain antibody fragments (ScFv) library was constructed against this parasite. Preliminary analysis indicated that a combined antibody library of about 2.08 X 10(5) individual clones and high diversity was generated. After four rounds of screening (bio-panning) against soluble spore protein prepared from lysed, intact, mature M rotundus spores, a strain monoclonal phage display ScFv, termed pCAN-6H9, with better affinity, was isolated. The pCAN-6H9 gene fragment was sequenced and analysed. The specificity of pCAN-6H9 was further demonstrated by dot-blot. In competition enzyme-linked immunosorbent assay, both the original and enriched phage-displayed ScFv repertoire showed significant inhibition of mouse anti-M rotundus serum binding to coated antigen, while the inhibition rate of monoclonal pCAN-6H9 phage particles was only 11.83%. |
英文摘要 | Immunological methods have been developed for the diagnosis of Myxobolus rotundus but their use has been limited for the prevention and therapy of this serious parasitic pathogen. Phage display antibody libraries are a powerful technique for the development of antibodies to molecules of interest and have advantages over traditional hybridroma approaches. In the present study, four antigen fractions related to M. rotundus were prepared and a combined phage display single-chain antibody fragments (ScFv) library was constructed against this parasite. Preliminary analysis indicated that a combined antibody library of about 2.08 X 10(5) individual clones and high diversity was generated. After four rounds of screening (bio-panning) against soluble spore protein prepared from lysed, intact, mature M rotundus spores, a strain monoclonal phage display ScFv, termed pCAN-6H9, with better affinity, was isolated. The pCAN-6H9 gene fragment was sequenced and analysed. The specificity of pCAN-6H9 was further demonstrated by dot-blot. In competition enzyme-linked immunosorbent assay, both the original and enriched phage-displayed ScFv repertoire showed significant inhibition of mouse anti-M rotundus serum binding to coated antigen, while the inhibition rate of monoclonal pCAN-6H9 phage particles was only 11.83%. |
学科主题 | Fisheries; Marine & Freshwater Biology; Veterinary Sciences |
WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
类目[WOS] | Fisheries ; Marine & Freshwater Biology ; Veterinary Sciences |
研究领域[WOS] | Fisheries ; Marine & Freshwater Biology ; Veterinary Sciences |
关键词[WOS] | MONOCLONAL-ANTIBODIES ; MYXOSPOREA ; MYXOZOA ; KNOWLEDGE |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000234976700001 |
公开日期 | 2010-10-13 |
内容类型 | 期刊论文 |
源URL | [http://ir.ihb.ac.cn/handle/152342/9044] ![]() |
专题 | 水生生物研究所_中科院水生所知识产出(2009年前)_期刊论文 |
作者单位 | 1.Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China 3.First Mil Med Univ, Inst Trop Med, Guangzhou, Guangdong, Peoples R China |
推荐引用方式 GB/T 7714 | Zhang, JY,Wang, JG,Wu, YS,et al. A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China[J]. JOURNAL OF FISH DISEASES,2006,29(1):1-7. |
APA | Zhang, JY,Wang, JG,Wu, YS,Li, M,Li, AH,&Gong, XL.(2006).A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China.JOURNAL OF FISH DISEASES,29(1),1-7. |
MLA | Zhang, JY,et al."A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China".JOURNAL OF FISH DISEASES 29.1(2006):1-7. |
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