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Although horse-derived antivenin is a major treatment, relatively strong and detrimental side effects are seen occasionally. In our study, yolk immunoglobulin (IgY) was purified from eggs, and DA protein was recognized using Western blotting and an enzyme-linked immunosorbent assay (ELISA), similar to therapeutic horse antivenin. The ELISA also indicated that specific IgY antibodies were elicited after the fifth booster, plateaued, and lasted for at least 3 months. To generate monoclonal single-chain variable fragment (scFv) antibodies, we used phage display technology to construct two libraries with short or long linkers, containing 6.24 \u00d7 10<jats:sup>8<\/jats:sup>and 5.28 \u00d7 10<jats:sup>8<\/jats:sup>transformants, respectively. After four rounds of biopanning, the eluted phage titer increased, and the phage-based ELISA indicated that the specific clones were enriched. Nucleotide sequences of 30 individual clones expressing scFv were analyzed and classified into four groups that all specifically recognized the DA venom protein. Furthermore, based on mass spectrometry, the scFv-bound protein was deduced to be snake venom metalloproteinase proteins. Most importantly, both IgY and mixed scFv inhibited the lethal effect in mice injected with the minimum lethal dosage of the DA protein. We suggest that together, these antibodies could be applied to the development of diagnostic agents or treatments for snakebite envenomation in the future.<\/jats:p>","DOI":"10.1128\/aem.02608-15","type":"journal-article","created":{"date-parts":[[2015,10,17]],"date-time":"2015-10-17T03:22:44Z","timestamp":1445052164000},"page":"71-80","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":28,"title":["Antibodies against Venom of the Snake Deinagkistrodon acutus"],"prefix":"10.1128","volume":"82","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3412-7400","authenticated-orcid":false,"given":"Chi-Hsin","family":"Lee","sequence":"first","affiliation":[{"name":"Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu-Ching","family":"Lee","sequence":"additional","affiliation":[{"name":"The Center of Translational Medicine, Taipei Medical University, Taipei, Taiwan"},{"name":"Ph.D. Program for Biotechnology in Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan"},{"name":"Core Facility for Antibody Generation and Research, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng-Huei","family":"Liang","sequence":"additional","affiliation":[{"name":"Core Facility for Antibody Generation and Research, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sy-Jye","family":"Leu","sequence":"additional","affiliation":[{"name":"Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan"},{"name":"Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liang-Tzung","family":"Lin","sequence":"additional","affiliation":[{"name":"Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan"},{"name":"Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jen-Ron","family":"Chiang","sequence":"additional","affiliation":[{"name":"Center for Research, Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi-Yuan","family":"Yang","sequence":"additional","affiliation":[{"name":"Core Facility for Antibody Generation and Research, Taipei Medical University, Taipei, Taiwan"},{"name":"School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan"},{"name":"Department of Laboratory Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2011.01.006"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(01)00232-X"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.3109\/15569549709016453"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0mt00017e"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4615-3806-6_15"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4613-0361-9_3"},{"key":"e_1_3_3_8_2","first-page":"781","article-title":"Characterization of the purified principles of Formosan snake venoms which affect blood coagulation and platelet aggregation","volume":"81","author":"Ouyang C","year":"1982","unstructured":"Ouyang C, Teng CM, Huang TF. 1982. 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