	US 12,391,765 B2
	Methods and materials for assessing response to plasmablast- and plasma cell-depleting therapies
Glennda Smithson, Antioch, IL (US); Jose Estevam, Quincy, MA (US); and Nicholas Jones, LaVergne, TN (US)
Assigned to Takeda Pharmaceutical Company Limited, Osaka (JP)
Filed by Takeda Pharmaceutical Company Limited, Osaka (JP)
Filed on Feb. 15, 2023, as Appl. No. 18/169,767.
Application 18/169,767 is a division of application No. 16/317,606, granted, now 11,613,586, previously published as PCT/US2017/042128, filed on Jul. 14, 2017.
Claims priority of provisional application 62/362,963, filed on Jul. 15, 2016.
Prior Publication US 2023/0357426 A1, Nov. 9, 2023
Int. Cl. C07K 16/28 (2006.01); A61K 39/00 (2006.01); A61P 19/02 (2006.01); G01N 33/569 (2006.01)

CPC C07K 16/2896 (2013.01) [A61P 19/02 (2018.01); G01N 33/56966 (2013.01); A61K 2039/505 (2013.01); C07K 2317/21 (2013.01); C07K 2317/732 (2013.01); C07K 2317/734 (2013.01); G01N 2333/91148 (2013.01)]

19 Claims

1. A method for assaying CD38-expressing cells in whole blood, the method comprising:

a) obtaining a sample of whole blood from a subject with an autoimmune disease, wherein the sample includes red blood cells, white blood cells and an anticoagulant;

b) lysing the red blood cells to form a treated sample;

c) freezing the treated sample to form a frozen sample, wherein freezing occurs within about 24 hours of obtaining the sample of whole blood from the subject;

d) measuring the level of CD38-expressing cells in the sample relative to a control subject without the autoimmune disease by flow cytometry within about 72 hours of obtaining the sample, wherein the frozen sample is warmed to room temperature before measuring the level of CD38-expressing cells, wherein the CD38-expressing cells are stained with an anti-CD38 antibody conjugated to a fluorochrome, and wherein the anti-CD38 antibody comprises:

1) a heavy chain variable region comprising:

i) a first CDR comprising SEQ ID NO: 3;

ii) a second CDR comprising SEQ ID NO: 4; and

iii) a third CDR comprising SEQ ID NO: 5; and

2) a light chain variable region comprising:

i) a first CDR comprising SEQ ID NO: 6;

ii) a second CDR comprising SEQ ID NO: 7; and

iii) a third CDR comprising SEQ ID NO: 8; or

1) a heavy chain variable region comprising:

i) a first CDR comprising SEQ ID NO: 13;

ii) a second CDR comprising SEQ ID NO: 14; and

iii) a third CDR comprising SEQ ID NO: 15; and

2) a light chain variable region comprising:

i) a first CDR comprising SEQ ID NO: 16;

ii) a second CDR comprising SEQ ID NO: 17; and

iii) a third CDR comprising SEQ ID NO: 18;

e) measuring the level of CD38-expressing cells relative to the control subject by assaying for free Ig light chains; and

f) measuring the expression of a biomarker for an elevated expression level of at least one CD38-expressing cell-enriched gene relative to the control subject, wherein the biomarker comprises mRNA and the at least one CD38-expressing cell-enriched gene comprises CD38.