title: [Screening of anti-human death receptor 5 antibody from a large natural phage antibody library].
abstract: OBJECTIVE
  To clone anti-human death receptor 5 (DR5) single-chain variable fragments (scFvs) from a large natural phage antibody library and identify the antigen-binding activity.
  
  
  METHODS
  Phage antibodies against human DR5 were screened from a large natural phage antibody library constructed in our laboratory by four rounds of panning process using eukaryotically expressed DR5-Fc protein. The antigen-binding activity of the obtained scFvs was determined by immunocytochemistry and ELISA, and then the scFVs were analyzed by DNA fingerprinting and sequence analysis.
  
  
  RESULTS
  After 4 rounds of panning, we obtained 26 positive clones with specific binding ability to DR5 protein. DNA fingerprinting and sequence analysis showed that there were four different variable fragments that were derived from different gene families. Soluble scFv fragments were prepared and then confirmed to bind specifically to DR5 protein in SW480 cells by immunocytochemistry.
  
  
  CONCLUSIONS
  Specific anti-human DR5 scFvs has been obtained from a large phage antibody library.
authors.affiliation: Department of General Surgery, Clinical College of Navy Medicine, Second Military Medical University, Navy General Hospital, Beijing 100048, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Jiang
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Lidong
authors.initials: L
authors.lastname: He
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Yuxiao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Li
authors.initials: L
authors.lastname: Ding
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Chenglin
authors.initials: C
authors.lastname: Wu
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Xiong
authors.affiliation: Central Laboratory, Navy General Hospital, Beijing 100048, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, Clinical College of Navy Medicine, Second Military Medical University, Navy General Hospital, Beijing 100048, China.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Wang
mesh_terms: D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D006801:Humans; D019151:Peptide Library; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012995:Solubility
chemical_list: D019151:Peptide Library; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D057127:Single-Chain Antibodies