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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; Wang, JG, Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China
2006
Source PublicationJOURNAL OF FISH DISEASES
ISSN0140-7775
Volume29Issue:1Pages:1-7
AbstractImmunological 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%.; 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%.
SubtypeArticle
KeywordCarassius Auratus China Combined Antibody Library Myxobolus Rotundus Phage Display Single-chain Antibody Fragments
DepartmentChinese 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
Subject AreaFisheries ; Marine & Freshwater Biology ; Veterinary Sciences
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaFisheries ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000234976700001
WOS KeywordMONOCLONAL-ANTIBODIES ; MYXOSPOREA ; MYXOZOA ; KNOWLEDGE
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9044
Collection期刊论文
Corresponding AuthorWang, JG, Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China
Affiliation1.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
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.,...&Wang, JG, Chinese Acad Sci, Hlth Cultivat Aquat Anim, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China.(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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