Methods are provided for directly converting histopathologically processed biological samples, tissues, and cells into a multiuse biomolecule lysate. This method allows for simultaneous extraction, isolation, solubilization, and storage of all biomolecules contained within the histopathologically processed biological sample, thereby forming a representative library of said sample. This multi-use biomolecule lysate is dilutable, soluble, capable of being fractionated, and used in biochemical assays.
What is claimed is:
1. A method of preparing a multi-use biomolecule lysate, comprising the steps of:
(a) heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature and a time sufficient to negatively affect protein cross-linking in said biological sample, wherein the temperature is at least 80° C. and the period of time is up to 24 hours; and
(b) treating the resulting composition with an effective amount of one or more proteolytic enzymes for a time sufficient to disrupt the tissue and cellular structure of said biological sample and to liquefy said sample, thereby producing a liquid, soluble, dilutable biomolecule lysate wherein the protein content of said lysate is representative of the total protein content of said histopathologically processed biological sample.
2. The method according to claim 1, wherein said histopathologically processed biological sample comprises a substantially homogeneous population of tissues or cells.
3. The method according to claim 1, further comprising, prior to step (a), the step of removing any paraffin present in said histopathologically processed biological sample by one or more methods selected from the group consisting of: adding an organic solvent;
heating; heating and adding a buffer comprising Tris; and heating and adding an organic solvent.
4. The method according to claim 1, further comprising prior to step (a) the step of mechanically disrupting said biological sample by at least one technique selected from the group consisting of: manual homogenization; vortexing; and physical mixing.
5. The method according to claim 1, wherein said biological sample is heated to a temperature between about 80° C. and about 100° C.
6. The method according to claim 1, wherein said biological sample is heated for a period of time from about 10 minutes to about 4 hours.
7. The method according to claim 1, wherein said proteolytic enzyme treatment is carried out at a temperature between about 37° C. to about 65° C.
8. The method according to claim 1 wherein said reaction buffer comprises a detergent.
9. The method according to claim 1 further comprising the step of fractionating said multi-use biomolecule lysate into distinct and separate biomolecule fractions.