Solid-phase monoclonal antibodies. A novel method of directing the function of biologically active molecules by presenting a specific orientation.
Monoclonal antibodies were produced against fibronectin and vitronectin, using the extracellular matrix from cultured bovine corneal endothelial cells as immunogen. Some antibodies inhibited the adhesion of BHK-21 cells to these molecules, in a standard assay with fibronectin or vitronectin coated on a plastic surface. By first coating the plastic surface with the monoclonal antibodies and subsequently binding vitronectin, the inhibitory effect was markedly increased. Divalent fibronectin required the presence of antibody in the medium as well as coated on the surface for maximal inhibition of cell adhesion. The inhibitory effect of surface-coated antibodies was stable in the presence of cells up to at least 24 h in vitro. Using this antibody coating method the surface could either be made compatible for cell adhesion (using non-inhibitory antibodies) or refractory. Assays performed by this method have considerable potential for quantifying particular functions of extracellular matrix molecules.
CSIRO Division of Biotechnology, Laboratory for Molecular Biology, North Ryde, NSW, Australia.
P A
PA
Underwood
J G
JG
Steele
B A
BA
Dalton
F A
FA
Bennett
D000818:Animals; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D002417:Cattle; D002448:Cell Adhesion; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000502:physiology; D005353:Fibronectins / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D011487:Protein Conformation; D019096:Vitronectin
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D006023:Glycoproteins; D019096:Vitronectin