Boston University

Chiron Diagnostics Corporation

Se-Kyung Oh

Harry Towbin

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.

Inhibition of protease activity of human whole blood cell lysates

A process for detecting cancer in a human patient is provided wherein a blood serum is recovered from the patient and is reacted with a monoclonal antibody comprising anti-haptoglobin variant under conditions to effect an immunoreaction when an immunosuppressive factor having a molecular weight of about 50K Daltons is present in the serum. The presence of immunosuppressive factor at concentration levels substantially exceeding normal concentration levels indicates the existence of immunoincompetence due to widely spread cancer. The effectiveness of cancer therapy is monitored by monitoring the change in levels of the immunosuppressive haptoglobin variant factor in the serum from the patient being treated.

Process for detecting cancer and for monitoring the effectiveness of

A high purity immunosuppression factor (IF) having a molecular weight of about 50,000 or a reduced IF having a molecular weight of about 25,000 derived from the higher molecular weight IF and which is stable in dilute aqueous solutions is obtained from human ascites fluid. An extract of human ascites fluid is subjected to chromatography to recover a component of the ascites fluid rich in the immunosuppression factor and containing proteinaceous material which does not degrade the IF factor. The pure IF is obtained by subjecting the component rich in IF to contact with solid phase anti-human IgG (Fc specific) to recover pure IF having a molecular weight of above 50,000. The reduced IF is obtained by heating the higher molecular weight IF.

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Brookline, MA, United States of America

Se-Kyung Oh