Inhibition of protease activity of human whole blood cell lysate

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 79, 435 792, 435 794, 435962, 435967, 435 7021, 43524027, 435 22, 435 23, 435 24, 435183, 435184, 435219, G01N 33567

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058637427

ABSTRACT:
A method of inhibiting proteolytic degradation of a thermally-stable intracellular protein is described. The method involves adding 1 or more denaturing agents to a sample which contains the protease and the protein of interest and heating the resulting solution at a temperature and for period of time sufficient to denature the protease. The method optionally includes a step for lysing the cell if the protein of interest is contained in a cell in order to release said protein. Additionally, a method of determining Mx protein induced by interferon in a blood sample is described. The method involves adding to a blood sample a lysing agent, a denaturing agent, and a detergent selected to solubilize Mx protein. The sample containing Mx protein is then heated at a temperature of from about 50.degree. C. to about 60.degree. C. for a period of time of from about 1 minute to about 30 minutes, and the Mx protein in the solution then is determined. Also described is a solution including a synthetic matrix mimicking whole blood cell lysates, or actual whole blood cell lysates, to which a known concentration of a thermally-stable intracellular protein is added to prepare a control material. In addition, a solution free of a protease that degrades the intracellular protein is disclosed, such solution remaining stable at 4.degree. C. for at least three weeks.

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