US 11,754,573 B2
Methods for the quantitation of polypeptides
Christopher Morgan, Southborough, MA (US); and Xiaokui Zhang, Northborough, MA (US)
Assigned to Genzyme Corporation, Cambridge, MA (US)
Filed by Genzyme Corporation, Cambridge, MA (US)
Filed on Jun. 16, 2021, as Appl. No. 17/349,840.
Application 17/349,840 is a continuation of application No. 16/246,376, filed on Jan. 11, 2019, granted, now 11,067,584.
Claims priority of provisional application 62/617,080, filed on Jan. 12, 2018.
Prior Publication US 2022/0003780 A1, Jan. 6, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 7/00 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/6857 (2013.01) [G01N 33/6848 (2013.01); G01N 33/6854 (2013.01)] 21 Claims
 
1. A method for quantitating an amount of a therapeutic polypeptide comprising a portion of an antibody heavy chain constant region in a sample comprising:
(a) digesting the sample comprising the therapeutic polypeptide comprising the portion of the antibody heavy chain constant region, wherein the portion of the antibody heavy chain constant region comprises an engineered mutation, and wherein digestion produces a peptide fragment derived from the antibody heavy chain constant region that is between 5 and 26 amino acids long and comprises the engineered mutation,
(b) analyzing the digested sample by mass spectrometry to determine quantity of the therapeutic peptide fragment, thereby determining the quantity of the therapeutic polypeptide comprising the portion of the antibody heavy chain constant region in the sample;
wherein the therapeutic polypeptide binds to an antigen selected from the group consisting of: A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H4/VTCN1, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2/MCP-1, CCL3/MIP-1a, CCL4/MIP-1b, CCL5/RANTES, CCL7/MCP-3, CCL8/mcp-2, CCL11/eotaxin, CCL15/MIP-1d, CCL17/TARC, CCL19/MIP-3b, CCL20/MIP-3a, CCL21/MIP-2, CCL24/MPIF-2/eotaxin-2, CCL25/TECK, CCL26/eotaxin-3, CCR3, CCR4, CD3, CD19, CD20, CD23/FCER2, CD24, CD27, CD28, CD38, CD39, CD40, CD70, CD80/B7-1, CD86/B7-2, CD122, CD137/41BB, CD137L, CD152/CTLA4, CD154/CD40L, CD160, CD272, CD273/PDL2, CD274/PDL1, CD275/B7H2, CD276/B7H3, CD278/ICOS, CD279/PD-1, CDH1/E-cadherin, chitinase, CLEC9, CLEC91, CRTH2, CSF-1/M-CSF, CSF-2/GM-CSF, CSF-3/GCSF, CX3CL1/SCYD1, CXCL12/SDF1, CXCL13, CXCR3, DNGR-1, ectonucleoside triphosphate diphosphohydrolase 1, EGFR, ENTPD1, FCER1A, FCER1, FLAP, FOLH1, Gi24, GITR, GITRL, GM-CSF, Her2, HHLA2, HMGB1, HVEM, ICOSLG, IDO, IFNα, IgE, IGF1R, IL2Rbeta, IL1, IL1A, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhIL10, IL12, IL13, IL13Ra1, IL13Ra2, IL15, IL17, IL17Rb/IL25, IL18, IL22, IL23, IL25, IL27, IL33, IL35, ITGB4/b4 integrin, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, NCR3LG1, NKG2D, NTPDase-1, OX40, OX40L, PD-1H, platelet receptor, PROM1, S152, SISP1, SLC, SPG64, ST2/receptor for IL33, STEAP2, Syk kinase, TACI, TDO, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP/IL7Ra, TSLPR, TWEAK, VEGF, VISTA, Vstm3, WUCAM, or XCR/GPR5/CCXCR1.