US 12,091,776 B2
Protease activity profiling via programmable phage display of comprehensive proteome-scale peptide libraries
Harry B. Larman, Baltimore, MD (US); Gabriel Roman-Melendez, Towson, MD (US); and Thiagarajan Venkataraman, Ellicott City, MD (US)
Assigned to The John Hopkins University, Baltimore, MD (US)
Filed by THE JOHNS HOPKINS UNIVERSITY, Baltimore, MD (US)
Filed on Jun. 26, 2020, as Appl. No. 16/913,572.
Claims priority of provisional application 62/867,319, filed on Jun. 27, 2019.
Prior Publication US 2020/0407467 A1, Dec. 31, 2020
Int. Cl. C40B 40/08 (2006.01); C07K 17/06 (2006.01); C12N 7/00 (2006.01); C12N 15/63 (2006.01); C12Q 1/6869 (2018.01)
CPC C40B 40/08 (2013.01) [C07K 17/06 (2013.01); C12N 7/00 (2013.01); C12N 15/63 (2013.01); C12Q 1/6869 (2013.01); C07K 2317/34 (2013.01); C07K 2319/43 (2013.01)] 33 Claims
OG exemplary drawing
 
1. A bacteriophage display vector comprising a nucleic acid sequence encoding:
(a) a peptide to be displayed on the surface of the bacteriophage, wherein the peptide is a candidate protease substrate encoded by a synthetic oligonucleotide;
(b) a first affinity tag distal to the peptide that immobilizes the bacteriophage to a solid support; and
(c) a second affinity tag proximal to the peptide for recapture of released bacteriophage following cleavage by a protease.