QUANTITATIVE DETERMINATION METHOD OF ERYTHROPOIETIN

• Main Authors: IMAI NOBUO, IGAWA YOUJI
• Format: Patent
• Language: English
• Published: 31.07.1984
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROCESSES USING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

PURPOSE:To determine quantitatively the amt. of erythropoietin (EPO) in the patient's bodily fluid at high sensitivity and in low cost with a simple operation by using the differentiation of the cell strain Cl-200 originating in the leukemic virus which is the friend of an anemic disease strain to a red blood cell system. CONSTITUTION:A sample contg. EPO is added to a medium wherein Cl-200 is contained preferably at about 5X10 -5X10 cells/ml and is cultured for 4-5 days preferably at 37 deg.C in an incubator contg. carbon dioxide at about 5%. 0.1ml benzidine soln. is added to 1.0ml medium upon completion of the culture, and after 10min, the dyed positive cells, i.e., the cells differentiated to the red blood cell system are counted on blood cell calculating plate and the ratio to the total number of cells is determined. The above-mentioned culture and dyeing are performed by using separately a sample contg. a known concn. of EPO and the ratios of the positive cells at each concn. of EPO are plotted on a logarithmic graph to make a calibration curve. The unknown concn. of EPO in the sample is determined from said calibration curve.