Monoclonal Antibody to Human Epidermal Growth Factor

Abstract

A monoclonal antibody directed to a species-specific determinant of human epidermal growth factor (h-EGF) was obtained by fusing murine myeloma cells with BALB/c mouse splenocytes sensitized to h-EGF. This antibody, referred to as 863.D4, did not react with either rat or mouse epidermal growth factor or with 11 other polypeptide hormones tested as shown by solid-phase radioimmunoassay (SPRIA), and immunoprecipitation followed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Scatchard analysis of the antibody binding to purified h-EGF revealed an apparent equilibrium dissociation constant of 1 x 10^-8M. The antibody blocked both the binding of h-EGF and h-EGF stimulation of ³H-thymidine incorporation into DNA by > 90% in confluent cultures of human foreskin fibroblasts.