ESTABLISHMENT OF AN ENZYME-IMMUNOASSAY SYSTEM FOR GLIOSTATIN PLATELET-DERIVED ENDOTHELIAL-CELL GROWTH-FACTOR (PD-ECGF)

A two-site enzyme immunoassay for gliostatin (GLS)/platelet-derived en dothelial cell growth factor (PD-ECGF) has been developed. The detecti on limit of gliostatin/PD-ECGF was 30 pg/well, and the optimal assay r ange was 0.1 to 10 ng/well. This assay system enabled us to confirm th e immunochemical identity of both factors and to detect immunoreactive gliostatin/PD-ECGF (IR-GLS/PD-ECGF) in human biological body fluids. The age-related analysis from newborn to 69 years revealed that the se rum IR-GLS/PD-ECGF level was high in infants younger than 1 year old ( 1.8 ng/ml) and in the 20-year-old age group (1.8 ng/ml), and highest i n the umbilical cord blood (2.1 ng/ml). Curiously high concentrations were detected in saliva with a significant sex difference (11.3 ng/ml for males and 48.7 ng/ml for females), and in synovial fluids (3.7 ng/ ml). A number of human tumor cells, gastric cancer cells, MKN-74, neur oblastoma cells, GOTO, as well as epidermoid carcinoma cells, A431, we re found to produce a significant amount of IR-GLS/PD-ECGF (0.2 to 21. 8 ng/mg protein), and some of them secreted the IR-GLS/PD-ECGF in the conditioned medium (approximately 0.5 ng/ml). The enzyme immunoassay s ystem is sufficiently sensitive for the basic and clinical study of gl iostatin/PD-ECGF in human body fluids. tissues and organs.