US 12,258,409 B2
Method of assessing the efficacy of a test agent for modulating programmed death 1 (PD-1) signaling by using anti-phosphotyrosinylated PD-1 monoclonal antibodies
Gordon J. Freeman, Brookline, MA (US); Vassiliki A. Boussiotis, Brookline, MA (US); Xia Bu, Brookline, MA (US); Vikram R. Juneja, Boston, MA (US); Arlene H. Sharpe, Brookline, MA (US); Nikolaos Patsoukis, Boston, MA (US); Jessica Weaver, Brighton, MA (US); and Laura Strauss, Boston, MA (US)
Assigned to Dana-Farber Cancer Institute, Inc., Boston, MA (US); Beth Israel Deaconess Medical Center, Boston, MA (US); and President and Fellows of Harvard College, Cambridge, MA (US)
Filed by Dana-Farber Cancer Institute, Inc., Boston, MA (US); Beth Israel Deaconess Medical Center, Boston, MA (US); and President and Fellows of Harvard College, Cambridge, MA (US)
Filed on Sep. 22, 2022, as Appl. No. 17/934,441.
Application 17/934,441 is a division of application No. 16/603,894, granted, now 11,492,403, previously published as PCT/US2018/028581, filed on Apr. 20, 2018.
Claims priority of provisional application 62/487,808, filed on Apr. 20, 2017.
Prior Publication US 2023/0331847 A1, Oct. 19, 2023
Int. Cl. C07K 16/44 (2006.01); C07K 16/28 (2006.01); G01N 33/50 (2006.01); G01N 33/574 (2006.01)
CPC C07K 16/2818 (2013.01) [C07K 16/44 (2013.01); G01N 33/5011 (2013.01); G01N 33/57492 (2013.01); C07K 2317/33 (2013.01); C07K 2317/34 (2013.01); C07K 2317/76 (2013.01); C07K 2317/92 (2013.01); G01N 2333/70596 (2013.01); G01N 2440/14 (2013.01); G01N 2500/04 (2013.01); G01N 2500/10 (2013.01); G01N 2500/20 (2013.01); G01N 2800/52 (2013.01)] 18 Claims
 
1. A method of assessing the efficacy of a test agent for modulating PD-1 signaling, the method comprising:
a) contacting a cell expressing PD-1 with a test agent; and
b) determining the ability of the test agent to modulate the level of phosphorylated PD-1 using at least one monoclonal antibody, or antigen-binding fragment thereof, that binds to a phosphorylation site at tyrosine residue 248 of human PD-1, wherein the monoclonal, or antigen-binding fragment thereof, comprises:
i) a light chain comprising CDR-L1 having the sequence of SEQ ID NO: 6, CDR-L2 having the sequence of SEQ ID NO: 9, and CDR-L3 having the sequence of SEQ ID NO: 12, and a heavy chain comprising CDR-H1 having the sequence of SEQ ID NO: 19, CDR-H2 having the sequence of SEQ ID NO: 22, and CDR-H3 having the sequence of SEQ ID NO: 25;
ii) a light chain comprising CDR-L1 having the sequence of SEQ ID NO: 34, CDR-L2 having the sequence of SEQ ID NO: 38, and CDR-L3 having the sequence of SEQ ID NO: 43, and a heavy chain comprising CDR-H1 having the sequence of SEQ ID NO: 52, CDR-H2 having the sequence of SEQ ID NO: 56, and CDR-H3 having the sequence of SEQ ID NO: 60;
iii) a light chain comprising CDR-L1 having the sequence of SEQ ID NO: 70, CDR-L2 having the sequence of SEQ ID NO: 74, and CDR-L3 having the sequence of SEQ ID NO: 78, and a heavy chain comprising CDR-H1 having the sequence of SEQ ID NO: 88, CDR-H2 having the sequence of SEQ ID NO: 92, and CDR-H3 having the sequence of SEQ ID NO: 96; or
iv) a light chain comprising CDR-L1 having the sequence of SEQ ID NO: 106, CDR-L2 having the sequence of SEQ ID NO: 110, and CDR-L3 having the sequence of SEQ ID NO: 114, and a heavy chain comprising CDR-H1 having the sequence of SEQ ID NO: 124, CDR-H2 having the sequence of SEQ ID NO: 128, and CDR-H3 having the sequence of SEQ ID NO: 132;
wherein a modulated level of phosphorylated PD-1 resulting from contacting with the test agent identifies the test agent as a modulator of PD-1 signaling.