A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China

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	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
	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.