US Patent:
20110275077, Nov 10, 2011
Inventors:
William James - Potomac MD, US
Vladimir Rait - Rockville MD, US
International Classification:
C12Q 1/68
C08G 73/10
C08G 64/42
C07H 21/00
C08G 63/91
US Classification:
435 611, 536 221, 536 254, 525 542
Abstract:
A method of analyzing tissue sections in a manner that provides information about the presence and expression levels of multiple biomarkers at each location within the tissue section. The method comprises the preparation of membranes having covalently bound oligonucleotides and the use of those membranes for evaluation of various markers in the sample. The membranes may be arranged in stacks, wherein each layer has a different oligonucleotide capture strand. Transfer oligonucleotides complementary to the capture strands are attached through a cleavable bond to antibodies that recognize and bind to specific biomarkers present in the tissue sample. The tissue sample is exposed to the antibody-transfer strand conjugate and then treated with a cleaving reagent. Upon cleavage, the transfer strand migrates through the stack and binds to the capture strand. The level of expression of the biomarker may be determined by measuring expression of a reporter on the transfer strand.