Human antibody fragments specific for the epidermal growth factor receptor selected from large non-immunised phage display libraries.
Antibodies to EGFR have been shown to display anti-tumour effects mediated in part by inhibition of cellular proliferation and angiogenesis, and by enhancement of apoptosis. Humanised antibodies are preferred for clinical use to reduce complications with HAMA and HAHA responses frequently seen with murine and chimaeric antibodies. We have used depletion and subtractive selection strategies on cells expressing the EGFR to sample two large antibody fragment phage display libraries for the presence of human antibodies which are specific for the EGFR. Four Fab fragments and six scFv fragments were identified, with affinities of up to 2.2nM as determined by BIAcore analysis using global fitting of the binding curves to obtain the individual rate constants (ka and kd). This overall approach offers a generic screening method for the identification of growth factor specific antibodies and antibody fragments from large expression libraries and has potential for the rapid development of new therapeutic and diagnostic reagents.
Souriau
Christelle
C
CSIRO Health Sciences and Nutrition, 343 Royal Parade, VIC. 3052, Australia.
Rothacker
Julie
J
Hoogenboom
Hennie R
HR
Nice
Edouard
E
D017209:Apoptosis; D001419:Bacteria / Q000378:metabolism; D001681:Biological Assay; D015374:Biosensing Techniques; D002460:Cell Line; D002469:Cell Separation; D002845:Chromatography; D004797:Enzyme-Linked Immunosorbent Assay; D066246:ErbB Receptors / Q000737:chemistry* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D007158:Immunologic Techniques; D007700:Kinetics; D019151:Peptide Library; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013997:Time Factors; D016211:Transforming Growth Factor alpha / Q000378:metabolism
D007128:Immunoglobulin Fragments; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D016211:Transforming Growth Factor alpha; D066246:ErbB Receptors