title: Construction of scFv library of human antibodies against tetanus toxin.
abstract: BACKGROUND
  Over the last three decades, monoclonal antibodies have become powerful therapeutics and diagnostics tools. Progress in the antibody engineering, and the appearance of great selection technology such as phage display has made human antibodies production possible against antigens with high affinities.
  
  
  OBJECTIVE
  The purpose of this study was to construct an immune antibody library from a vaccinated donor against tetanus toxin.
  
  
  METHODS
  A blood sample was drawn from the donor who was vaccinated with tetanus toxoid. PBMC were isolated by using ficoll. After RNA extraction and cDNA synthesis. In order to amplify VH and VL regions, two uniplex PCRs were performed and put considering NcoI & HindIII, MluI & NotI restriction sites respectively for their cloning. These amplicons were cloned to pSEX81 vector and transformed to E. coli XL1blue strain. Eventually, recombinant plasmids were extracted and sequenced.
  
  
  RESULTS
  The result showed acceptable similarity between antibody gene library nucleotide sequences and the antibody genes were deposited in this database.
  
  
  CONCLUSIONS
  In this study, the VH and VL genes human antibody library was constructed and confirmed by using DNA sequencing and alignment of sequences with blast database.
authors: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors: Hamideh Rouhani
authors: HR
authors: Nejad
authors: Department of Bioscience and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran.
authors: Jalil Fallah
authors: JF
authors: Mehrabadi
authors: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors: Jamile
authors: J
authors: Nourozi
authors: Department of Immunology, Pasteur Institute of Iran, Tehran, Iran.
authors: Mehdi
authors: M
authors: Mahdavi
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003017:Clostridium tetani / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D017422:Sequence Analysis, DNA; D061905:Single-Domain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D013742:Tetanus / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D013744:Tetanus Toxin / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology; D014611:Vaccination
chemical_list: D019151:Peptide Library; D011994:Recombinant Proteins; D061905:Single-Domain Antibodies; D013744:Tetanus Toxin
keywords: Antibody library gene; monoclonal antibody; tetanus toxin