title: [Isolation and identification of single chain Fv antibodies against breast cancer from a human phage display library].
abstract: OBJECTIVE
  To isolate and identify single chain Fv (scFv) antibodies against breast cancer from a constructed human phage display library.
  
  
  METHODS
  Recombinant phages specific for breast cancer cells were enriched after four-round screening with MCF-7 cells. We selected the antigen-positive ones from the enriched clones by phage ELISA. The positive clones were used to infect E.coli HB2151 to express soluble scFv antibody. The antigen binding activity of the soluble antibodies was detected by Western blotting.
  
  
  RESULTS
  The specific antibodies against MCF-7 cells were enriched after four rounds of affinity selection. SDS-PAGE and Western blotting showed a band at relative molecular mass 30 000 Da, which indicated soluble antibodies were present. ELISA analysis revealed that soluble antibodies had the affinity to a human breast cancer cell line MCF-7 but not to other cancer cell line, which demonstrated scFv could react specifically with breast cancer cells.
  
  
  CONCLUSIONS
  We constructed a scFv phage library against human breast cancer with high capacity by phage display technology. The scfv was demonstrated successfully to be expressed in E.coli HB215 and have a specificity to breast cancer. Our findings may provide an alternative approach, and a basis for further studies on diagnosis and therapy of breast cancer.
authors.affiliation: Department of Pathology and Pathophysiology, Fourth Military Medical University, Xi'an, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yuan
authors.forename: Yan
authors.initials: Y
authors.lastname: Cui
authors.forename: Jing
authors.initials: J
authors.lastname: Ye
authors.forename: Qing
authors.initials: Q
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D001483:Base Sequence; D001943:Breast Neoplasms / Q000276:immunology*; D061505:Cell Surface Display Techniques; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D008969:Molecular Sequence Data; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012995:Solubility
chemical_list: D019151:Peptide Library; D057127:Single-Chain Antibodies