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 endothelial cell growth factor (PD-ECGF) has been developed. The detection limit of gliostatin/PD-ECGF was 30 pg/well, and the optimal assay range was 0.1 to ng/well. This assay system enabled us to confirm the 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 serum 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 in 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, neuroblastoma cells, GOTO, as well as epidermoid carcinoma cells, A431, were 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 system is sufficiently sensitive for the basic and clinical study of gliostatin/PD-ECGF in human body fluids, tissues and organs.
Hirano
T
T
Department of Bioregulation Research, Nagoya City University Medical School, Japan.
Asai
K
K
Matsukawa
K
K
Kato
T
T
Takeuchi
M
M
Yonezawa
M
M
Otsuka
T
T
Matsui
N
N
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D002648:Child; D002675:Child, Preschool; D005260:Female; D005312:Fetal Blood / Q000737:chemistry; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000097:blood; D012463:Saliva / Q000737:chemistry; D012680:Sensitivity and Specificity; D012737:Sex Factors; D013582:Synovial Fluid / Q000737:chemistry; D013939:Thymidine Phosphorylase / Q000032:analysis*; D014407:Tumor Cells, Cultured / Q000378:metabolism
D009419:Nerve Tissue Proteins; D013939:Thymidine Phosphorylase