IP Library Granted Patent US 9,535,070
Granted Patent B2
US 9,535,070 · App. 14/073,332 · Granted Jan 3, 2017

High throughput detection of fusion proteins

Inventors: Justin Saul (Mesa, AZ); Ji Qiu (Chandler, AZ); Joshua LaBaer (Chandler, AZ); Mitch Magee (Chandler, AZ); John Chaput (Phoenix, AZ); Sujay Sau (Mesa, AZ)
Assignee: ARIZONA BOARD OF REGENTS, A BODY CORPORATE OF THE STATE OF ARIZONA, ACTING FOR AND ON BEHALF OF ARIZONA STATE UNIVERSITY
G01N33/582
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Quick Facts
Patent No.
US 9,535,070
App. No.
14/073,332
Granted
Jan 3, 2017
Kind
B2
Abstract

A method and related microfluidic chip and kit for high throughput detection of proteins of interest contained in a sample is disclosed. The method comprises of specifically labeling fusion proteins in a complex sample with fusion tag specific fluorophores that specifically bind the fusion tags coupled to the proteins of interest, and subjecting the sample to automated capillary electrophoresis, wherein the presence of the proteins of interest in the sample is detected by fluorescence signals associated with the fusion tag specific fluorophores.

Claims (7)

1. A method for high throughput detection of proteins of interest contained in a sample, comprising:

(a) contacting a sample not subjected to prior purification comprising contaminating proteins without fusion tags and affinity tagged fusion proteins of interest to a plurality of fluorescent labels having specificity for the affinity tags, wherein, upon contacting, fluorescent labels covalently bind the affinity tags to form detectable protein complexes;

(b) subjecting the contacted sample to microfluidic electrophoresis, wherein the presence of the proteins of interest but not contaminating proteins in the contacted sample is detected by fluorescence signals associated with the affinity tags.

2. The method of claim 1 , wherein the sample not subjected to prior purification is a cell lysate.

3. The method of claim 1 , wherein the sample not subjected to prior purification is a bacterial cell lysate.

4. The method of claim 1 , wherein the fusion tag is a fusion tag protein.

5. The method of claim 1 , wherein the fusion tag specific fluorophore has an excitation and emission maxima near about 635 nm and about 700 nm, respectively.

Assignments (3)
CONFIRMATORY LICENSE Recorded Oct 20, 2016
From: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 040436/0276 →
CONFIRMATORY LICENSE Recorded Oct 20, 2016
From: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 040436/0362 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 17, 2013
From: SAUL, JUSTIN; MAGEE, MITCH; QIU, JI; LABAER, JOSHUA; SAU, SUJAY; CHAPUT, JOHN
To: ARIZONA BOARD OF REGENTS, A BODY CORPORATE OF THE STATE OF ARIZONA, ACTING FOR AND ON BEHALF OF ARIZONA STATE UNIVERSITY
Reel/Frame 031798/0442 →
Continuity (2)
Provisional Application 61724608 · Nov 9, 2012
Related Publication 20140141991A1 · May 22, 2014