Isolation of tumor cell-specific single-chain Fv from immunized mice using phage-antibody libraries and the re-construction of whole antibodies from these antibody fragments.
Enhanced expression of epidermal growth factor receptor (EGFR) occurs on a variety of malignant tissues thus making anti-EGFR antibodies possible agents for the diagnosis and therapy of human tumors. Standard hybridoma technology has been used successfully to isolate anti-EGFR antibodies from immunized mice and rats. This report demonstrates that phage-antibody libraries are an alternative, and more versatile, method for isolating antibodies from immunized mice. Anti-EGFR antibodies were isolated from phage-antibody libraries constructed not only from the spleen of an immunized mouse but also from the draining lymph node of an immunized mouse and from in vitro immunized mouse cells. Two of the single-chain Fv isolated from the phage-antibody libraries were engineered to create partially humanized whole antibody molecules.
Kettleborough
C A
CA
MRC Collaborative Centre, Mill Hill, London.
Ansell
K H
KH
Allen
R W
RW
Rosell-Vives
E
E
Güssow
D H
DH
Bendig
M M
MM
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D001435:Bacteriophages / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D066246:ErbB Receptors / Q000276:immunology*; D015723:Gene Library; D006801:Humans; D007114:Immunization; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D051379:Mice; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv; D066246:ErbB Receptors