Liquid tissue preparation from histopathologically processed biological samples, tissues and cells
AssigneeExpression Pathology, Inc.
InventorsMarlene M. Darfler, David B. Krizman
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Abstract
The current invention provides a method for directly converting histopathologically processed biological samples, tissues, and cells into a multi-use biomolecule lysate. This method allows for simultaneous extraction, isolation, solublization, 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 any number of subsequent experiments.
Claims15
A method of preparing a biomolecule lysate, comprising the steps of:
- (a) heating a composition comprising a formalin fixed biological sample and a reaction buffer at a temperature from 80° C. to 100° C. for a period of time from 10 minutes to 4 hours to reverse or release protein cross-linking in said biological sample, and
- (b) treating the resulting composition with an effective amount of a proteolytic enzyme selected from the group consisting of trypsin, chymotrypsin, and endoproteinase Lys-C for a period of time from 30 minutes to 24 hours at a temperature from 37° C. to 65° C. to disrupt the tissue and cellular structure of said biological sample and to liquefy said sample, thereby producing a liquid, soluble, dilutable biomolecule lysate that is suitable for protein analysis and wherein the protein content of said lysate is representative of the total protein content of said biological sample.
A method of preparing a biomolecule lysate, comprising the steps of:
- (a) heating a composition comprising a formalin fixed biological sample and a reaction buffer at a temperature from 80° C. to 100° C. for a period of time from 10 minutes to 4 hours to reverse or release protein cross-linking in said biological sample,
- (b) treating the resulting composition with an effective amount of a proteolytic enzyme selected from the group consisting of trypsin, chymotrypsin, and endoproteinase Lys-C for a period of time from 30 minutes to 24 hours at a temperature between 37° C. to 65° C. to disrupt the tissue and cellular structure of said biological sample and to liquefy said sample, thereby producing a liquid, soluble, dilutable biomolecule lysate that is suitable for protein analysis and wherein the protein content of said lysate is representative of the total protein content of said biological sample, and
- fractionating said biomolecule lysate into distinct and separate biomolecule fractions.
Dependencies are inferred from the claim text.