Recombinant Human Monoclonal Fab Fragments against Rotaviras from Phage Display Combinatorial Libraries

• Itoh Kunihiko

  Department of Pharmaceutical Science, Akita University Hospital

• Nakagomi Osamu

  Department of Microbiology, Akita University School of Medicine

• Suzuki Kaori

  Department of Pharmaceutical Science, Akita University Hospital

• Inoue Kazuyuki

  Department of Pharmaceutical Science, Akita University Hospital

• Tada Hitoshi

  Department of Pharmaceutical Science, Akita University Hospital

• Suzuki Toshio

  Department of Pharmaceutical Science, Akita University Hospital

書誌事項

タイトル別名

• Recombinant Human Monoclonal Fab Fragments against Rotavirus from Phage Display Combinatorial Libraries.

抄録

We prepared and characterized human monoclonal Fab fragments to rotavirus from IgG₁ κ combinatorial libraries (designated as N and O) constructed from peripheral blood lymphocytes (PBLs) from two healthy individuals. Approximately 30-fold enrichment in eluted phage was obtained in these libraries after five rounds of panning against rabbit polyclonal antibody-captured human rotavirus (HRV) Wa strain. Forty-eight clones from each library. were tested for reactivity to HRV Wa in an enzyme-linked immunosorbent assay (ELISA), and the identities of positive clones were determined by BstNI fingerprinting. As a result, eight individual clones (five from N library and three from O library) were isolated. In testing the cross-reactivity of Fabs against a panel of self- or non-self antigens, all Fab clones were found to be specific for HRV Wa. Fab clones from the two libraries showed distinct characteristics with respect to their reaction patterns with Wa and crossreactivities with rotavirus strains, and displayed variable heavy (V_H) chain gene usage, although they recognized the VP6 protein as determined by immunoblotting. The distinct epitope recognition by Fabs from two libraries suggests different courses of humoral immune response to rotavirus during infection in the two individuals.

収録刊行物

• The Journal of Biochemistry

  The Journal of Biochemistry 125 (1), 123-129, 1999

  社団法人 日本生化学会

キーワード

• combinatorial library
• human antibody repertoire
• phage display
• recombinant Fab fragment
• rotavirus infection

詳細情報

• CRID: 1390282679907714944
• NII論文ID: 130003533597
• NII論文ID: 10005461965
• NII書誌ID: AA00694073
• DOI: 10.1093/oxfordjournals.jbchem.a022248
• COI: 1:CAS:528:DyaK1MXhs1yqt7s%3D
• ISSN: 17562651
• ISSN: 0021924X
• ISSN: http://id.crossref.org/issn/0021924X
• NDL書誌ID: 4639636
• PubMed: 9880807
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I4639636
• Web Site: http://academic.oup.com/jb/article-pdf/125/1/123/2583854/125-1-123.pdf
• Web Site: https://search.jamas.or.jp/link/ui/1999134326
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可