The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Apr. 21, 2020

Filed:

Mar. 25, 2015
Applicant:

British Columbia Cancer Agency, Vancouver, CA;

Inventors:

Robert Holt, Vancouver, CA;

Govinda Sharma, Vancouver, CA;

Assignee:

British Columbia Cancer Agency Branch, Vancouver, British Columbia, CA;

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 5/00 (2006.01); C12N 5/02 (2006.01); G01N 33/53 (2006.01); G01N 33/68 (2006.01); C12Q 1/6886 (2018.01); C12N 15/10 (2006.01); G01N 33/50 (2006.01); G01N 33/569 (2006.01);
U.S. Cl.
CPC ...
G01N 33/6878 (2013.01); C12N 15/1037 (2013.01); C12N 15/1086 (2013.01); C12Q 1/6886 (2013.01); G01N 33/5091 (2013.01); G01N 33/56972 (2013.01); G01N 33/6845 (2013.01); C12Q 2600/158 (2013.01);
Abstract

The present invention is a method for determining the identity of the epitopes recognized by T-cells. The method consists of expressing an encoded library of candidate epitope sequences in a recipient reporter cell capable of providing a detectable signal upon cytotoxic attack from a single cognate T-cell followed by contacting the reporter cells with T-cells of interest. The reporter cells with a single indicating cytotoxic attack from a T-cell are isolated and then analyzed by next-generation sequencing in order to identify the epitope sequences. Specifically disclosed is a method in which a library of candidate epitope-encoding nucleic acids are expressed in cells which feature a membrane-bound major histocompatibility complex (MHC) protein, said library produced by transfection of plasmids featuring both a nucleotide encoding the candidate epitope and a nucleotide encoding a FRET-based fluorescent protein cleaved by granzyme.


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