US 12,422,440 B2
Newborn screening for primary immunodeficiencies, cystinosis, and Wilson disease
Sihoun Hahn, Clyde Hill, WA (US); Sunhee Jung, Seattle, WA (US); Jeffrey Whiteaker, Seattle, WA (US); Troy Torgerson, Seattle, WA (US); Amanda Paulovich, Seattle, WA (US); Christopher Collins, Seattle, WA (US); and Remwilyn Dayuha, Lynnwood, WA (US)
Assigned to Seattle Children's Hospital, Seattle, WA (US); and Fred Hutchinson Cancer Research Center, Seattle, WA (US)
Appl. No. 17/282,989
Filed by Seattle Children's Hospital, Seattle, WA (US); and Fred Hutchinson Cancer Center, Seattle, WA (US)
PCT Filed Oct. 4, 2019, PCT No. PCT/US2019/054856
§ 371(c)(1), (2) Date Apr. 5, 2021,
PCT Pub. No. WO2020/072996, PCT Pub. Date Apr. 9, 2020.
Claims priority of provisional application 62/742,161, filed on Oct. 5, 2018.
Prior Publication US 2021/0341492 A1, Nov. 4, 2021
Int. Cl. G01N 33/68 (2006.01); C07K 16/28 (2006.01)
CPC G01N 33/6893 (2013.01) [C07K 16/2809 (2013.01); G01N 33/6848 (2013.01); C07K 2317/34 (2013.01); C07K 2317/565 (2013.01); G01N 2800/24 (2013.01); G01N 2800/60 (2013.01)] 16 Claims
 
1. A method of detecting signature peptides of severe combined immunodeficiency (SCID), Wiskott-Aldrich syndrome (WAS), and/or X-linked agammaglobulinemia (XLA) in a dried blood spot (DBS) sample, the method comprising: obtaining a DBS sample; digesting proteins from the DBS sample with an enzyme to yield a digested DBS sample comprising peptides; enriching for:
a CD3ε signature peptide of SCID having the sequence of SEQ ID NO: 1 with an antibody or antigen-binding fragment thereof that binds the CD3ε signature peptide and comprises: a heavy chain variable (VH) domain comprising a complementarity determining region (CDR) 1 of SEQ ID NO: 22, a CDR2 of SEQ ID NO: 23, and a CDR3 of SEQ ID NO: 24, and a light chain variable (VL) domain comprising a CDR1 of SEQ ID NO: 25, a CDR2 of SEQ ID NO: 26, and a CDR3 of SEQ ID NO: 27;
a first WASp signature peptide of WAS having the sequence of SEQ ID NO: 2 with an antibody or antigen-binding fragment thereof that binds the first WASp signature peptide and comprises: a VH domain comprising a CDR1 of SEQ ID NO: 28, a CDR2 of SEQ ID NO: 29, and a CDR3 of SEQ ID NO: 30, and a VL domain comprising a CDR1 of SEQ ID NO: 31, a CDR2 of SEQ ID NO: 32, and a CDR3 of SEQ ID NO: 33; and
a BTK signature peptide of XLA having the sequence of SEQ ID NO: 4 with an antibody or antigen-binding fragment thereof that binds the first BTK signature peptide and comprises: a VH domain comprising a CDR1 of SEQ ID NO: 34, a CDR2 of SEQ ID NO: 35, and a CDR3 of sequence GDI, and a VL domain comprising a CDR1 of SEQ ID NO: 36, a CDR2 of SEQ ID NO: 37, and a CDR3 of SEQ ID NO: 38; and performing liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM-MS) on the enriched peptides to determine a concentration for each signature peptide, thereby detecting signature peptides of SCID, WAS, and XLA in the DBS sample.