METHOD FOR QUICK QUANTITATIVE DETERMINATION OF LYOPHILIZED BCG VACCINE SPECIFIC ACTIVITY Russian patent published in 2017 - IPC G01N21/76 

FIELD: medicine.

SUBSTANCE: invention relates to a method for rapid quantification of the specific activity of a lyophilized BCG vaccine comprising lyophilized vaccine reconstruction, extracellular ATP removal using apyrase, intracellular ATP extraction, intracellular ATP content determination by a bioluminescence method, and calculation of specific activity. Lyophilized vaccine reconstruction is carried out in physiological solution, extracellular ATP removal carried out by incubating the reconstituted vaccine with apyrase, intracellular ATP extraction carried out by addition of DMSO, ATP content determination in the obtained extract is carried out according to the bioluminescence signal intensity for a mixture comprising the reconstituted BCG vaccine extract in DMSO and ATP-reagent solution based on luciferin-luciferase, intracellular ATP content calculation in a sample of reconstituted vaccines is performed, using the bioluminescent signal intensity for ATP-control, which is a lyophilized preparation with a known ATP content, calculation of the specific intracellular ATP content in the analyzed sample is performed taking into account the lyophilized vaccine weight in the sample to be analyzed, specific activity is then calculated by the corresponding formula.

EFFECT: analysis duration reduced to 2 hours, simplification and standardization of the procedure for lyophilized BCG vaccine reconstruction, reduced duration and laboriousness of the method for intracellular ATP extraction and bioluminescence signal measurement, unified composition of the reagents used in the analysis.

2 ex, 1 dwg, 3 tbl