Patricia Sammie Grom

US Patent:

20060046272, Mar 2, 2006

Filed:

Aug 27, 2004

Appl. No.:

10/928570

Inventors:

Sue Chow - Markham, CA
David Hedley - Toronto, CA
T. Shankey - Miami FL, US
Patricia Grom - Sunrise FL, US

International Classification:

G01N 33/567
G01N 33/53

US Classification:

435007200

Abstract:

This invention is directed to a method for preparation of a biological sample for measurement of protein epitopes that allows for the preservation of intracellular protein epitopes and detection of signal transduction pathways based on the ability to capture transient activation states of the epitopes. The method provided by the invention allows for the rapid fixation of biological samples containing red blood cells, to ensure that epitopes of signal transduction molecules and other intracellular protein epitopes are preserved in the active state. The method of the invention further allows for lysis of red blood cells, thereby making it a useful method for cytometric analysis of biological samples, including, for example, whole blood, bone marrow aspirates, peritoneal fluids, and other red blood cell containing samples. The invention also provides a method to recover or “unmask” epitopes on intracellular antigens that have been made inaccessible by the cross linking fixative necessary to fix the sample. Significantly, the methods of the invention allow preservation and analysis of phospho-epitope levels in biological samples taken directly from patients to determine disease-specific characteristics.