The EF-hand
calcium-binding protein S100B presents a wide range of biological activities and functions. This
binding protein is involved in various human diseases, including
cancer, brain trauma and
ischemia, neuro-degenerative disease (
Alzheimer's disease), and
psychiatric disorders. In this study, we prepared
human S100B protein and its
monoclonal antibodies.
Human S100B protein was expressed in Escherichia coli, successfully purified by
diethylaminoethyl cellulose anion-exchange chromatography, and then identified by western blot analysis.
Monoclonal antibodies (mAbs) were produced by the standard hybridoma method and validated by
enzyme-linked
immunosorbent assay and western blot analysis. The prepared
human S100B protein and its mAbs demonstrated potential biological activities. The KD of one mAb is approximately 4.72x10-8 mol/l, and its cross reactivity is low with human S100A4, mouse S100A4, and human S100A1. Recombinant Soluble S100B can promote the migration and invasion of HeLa cells. The expression of S100B
protein in
tumor tissues can be detected effectively by using the prepared
monoclonal antibodies. Increasing concentration of the anti-human S100B mAbs showed a reduced expression of the S100B
protein. Subsequently, the expression of p53 increased significantly (P<0.05) in A375 cells. A significant increase in apoptosis in A375 cells was observed with increasing S100B mAb concentration. Results showed that our prepared S100B mAbs were suitable for detecting S100B expression in human tissues, furnishing promising tools for further functional investigation and clinical applications.