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学科主题: Fisheries; Marine & Freshwater Biology; Veterinary Sciences
题名: A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China
作者: Zhang, JY; Wang, JG; Wu, YS; Li, M; Li, AH; Gong, XL
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
关键词: Carassius auratus ; China ; combined antibody library ; Myxobolus rotundus ; phage display ; single-chain antibody fragments
刊名: JOURNAL OF FISH DISEASES
发表日期: 2006
卷: 29, 期:1, 页:1-7
收录类别: SCI
文章类型: Article
部门归属: Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China; Chinese Acad Sci, Grad Sch, Beijing, Peoples R China; First Mil Med Univ, Inst Trop Med, Guangzhou, Guangdong, Peoples R China
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
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%.
关键词[WOS]: MONOCLONAL-ANTIBODIES ; MYXOSPOREA ; MYXOZOA ; KNOWLEDGE
语种: 英语
WOS记录号: WOS:000234976700001
ISSN号: 0140-7775
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/9044
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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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

Recommended Citation:
Zhang, JY; Wang, JG; Wu, YS; Li, M; Li, AH; Gong, XL.A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China,JOURNAL OF FISH DISEASES,2006,29(1):1-7
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