Method for stabilization of proteins in solution
The present invention relates to a method for stabilization of analytes in solutions of solubilized body samples. The method comprises the steps of solubilizing the body samples obtained from a subject in a suitable sample medium and stabilizing said body sample contained within the sample medium by heating said sample medium for a certain period of time. A further aspect of the invention is a method of a denaturing immunoassay of proteins in solution. The method comprises the steps of bringing a sample containing proteins into contact with a denaturing agent and heating said sample in the presence of the denaturing agent to allow the protein to be denatured.
1. A method for preparing and stabilizing one or more analytes in solutions for a denaturing immunoassay of body samples, comprising the steps of:
i) obtaining a solubilized mammalian body sample containing one or more analytes in a sample medium comprising a denaturing agent of SDS at 0.1-1% w/v, wherein the one or more analytes are selected from the group consisting of p16 INK4a , EpCAM, gCatenin, MCM2, MCM3, MCM4, MCM5, MCM6, and MCM7;
ii) subsequently heating said sample medium containing the one or more analytes to a temperature of 90-110 ±2° C. for a period of time of 3 to 45 minutes;
iii) stabilizing and storing the one or more heated analytes in said sample medium, and
iv) subsequently performing a denaturing immunoassay on said sample medium containing the one or more analytes and the denaturing agent of SDS.
2. The method according to claim 1 , wherein the temperature is between 95° C. and 99° C.
3. The method according to claim 1 , wherein the period of time is at least 5 minutes.
4. The method according claim 3 , wherein the period of time is between 7 minutes and 30 minutes.
5. The method according to claim 4 , wherein the period of time is between 10 minutes and 27 minutes.
6. The method of claim 5 , wherein the period of time is between 20 and 25 minutes.
7. The method according to claim 1 , wherein the heating step is performed in a heating device selected from the group consisting of a water bath, a pressure cooker, a microwave oven, an UV heating device, and a heating block.
8. The method according to claim 1 , wherein the body sample is selected from the group consisting of secretions, swabs, lavages, body fluids, semen, cell- and tissue-samples, liquid based cytology samples, blood, smears, sputum, urine, stool, Liquor cerebrospinalis, bile, gastrointestinal secretions, lymph, bone marrow, aspirates and biopsies of organs.
9. The method according to claim 1 , wherein the one or more analytes are p16 INK4a , EpCAM or gCatenin.
10. The method according to claim 1 , wherein said storing is carried out at 2-30° C.
11. The method according to claim 1 , wherein said storing is carried out for 1, 2, 5, or 6 days.
12. The method according to claim 1 , wherein said storing is carried out for up to 14 days.
13. The method according to claim 1 , wherein said sample medium of (iii) is stored without further handling until subsequently performing the denaturing immunoassay.
14. The method according to claim 1 , wherein said denaturing immunoassay is selected from the group consisting of immunoprecipitation assays, immunological assays, EIA, ELISA, RIA, lateral flow assays, flow through assays, immunochromatographic strips, and latex agglutination assays.
15. The method according to claim 9 , wherein the analyte is p16 INK4a .
16. The method according to claim 1 , wherein the analyte is MCM-2 or MCM-5.