Isolation and characterisation of vascular endothelial growth factor-165 specific scFv fragments by phage display.
Vascular endothelial growth factor (VEGF) is a multifunctional cytokine which plays a major role in angiogenesis. Alternative splicing causes the production of several different isoforms (VEGF-A 121, 145, 165, 189, 206). VEGF is essential for tumor angiogenesis and several studies have correlated elevated VEGF levels with tumor stage, metastases and progression. Antibody phage display was employed to isolate two scFv antibody fragments, D8 and F10, with specificity for the VEGF165 isoform. It was shown by ELISA and competitive immunohistochemistry that each clone bound to VEGF165 but not VEGF121. Immunohistochemistry with D8 and F10 on colorectal carcinoma and adenoma sections revealed positive staining similar to that shown by a polyclonal VEGF antibody. The scFv antibody fragments, D8 and F10, will be useful in specifically detecting circulating VEGF165 in cancer patients as most studies to date have quantified the total level of circulating VEGF (121 and 165). These reagents will allow further elucidation of the role of VEGF in tumor angiogenesis.
Smith
K A
KA
Department of Surgery, Medical Research Laboratory, Wolfson Building, University of Hull, Hull, HU6 7RX, UK.
Kirkpatrick
N
N
Madden
L A
LA
Topping
K P
KP
Monson
J R T
JR
Greenman
J
J
D000236:Adenoma / Q000737:chemistry / Q000473:pathology; D000595:Amino Acid Sequence; D000918:Antibody Specificity; D001667:Binding, Competitive; D014408:Biomarkers, Tumor / Q000032:analysis / Q000276:immunology; D002277:Carcinoma / Q000737:chemistry / Q000473:pathology; D015179:Colorectal Neoplasms / Q000737:chemistry / Q000473:pathology; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000276:immunology; D016228:Endothelial Growth Factors / Q000032:analysis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005629:Frozen Sections; D006801:Humans; D007124:Immunoenzyme Techniques; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D007163:Immunosorbent Techniques; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000276:immunology*; D008222:Lymphokines / Q000032:analysis / Q000276:immunology*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis / Q000276:immunology; D009389:Neovascularization, Pathologic / Q000175:diagnosis; D019151:Peptide Library; D016415:Sequence Alignment; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D014408:Biomarkers, Tumor; D022801:Complementarity Determining Regions; D016228:Endothelial Growth Factors; D007135:Immunoglobulin Variable Region; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D009363:Neoplasm Proteins; D019151:Peptide Library; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors