US 12,247,985 B2
Methods of detecting transthyretin
Joshua Reginald Salmans, South San Francisco, CA (US); Svetlana Alexander, Sunnyvale, CA (US); Robin Barbour, Walnut Creek, CA (US); Jianmin Li, Castro Valley, CA (US); Jeffrey N. Higaki, San Mateo, CA (US); and Tarlochan S. Nijjar, Orinda, CA (US)
Assigned to NOVO NORDISK A/S, Bagsvaerd (DK)
Appl. No. 16/753,661
Filed by Novo Nordisk A/S, Bagsvaerd (DK)
PCT Filed Oct. 5, 2018, PCT No. PCT/US2018/054723
§ 371(c)(1), (2) Date Apr. 3, 2020,
PCT Pub. No. WO2019/071206, PCT Pub. Date Apr. 11, 2019.
Claims priority of provisional application 62/569,438, filed on Oct. 6, 2017.
Claims priority of provisional application 62/579,817, filed on Oct. 31, 2017.
Claims priority of provisional application 62/647,582, filed on Mar. 23, 2018.
Prior Publication US 2020/0249244 A1, Aug. 6, 2020
Int. Cl. G01N 33/68 (2006.01); A61K 39/395 (2006.01); A61K 49/00 (2006.01); C07K 16/18 (2006.01); G01N 21/66 (2006.01); G01N 21/76 (2006.01); G01N 33/543 (2006.01); A61K 39/00 (2006.01)
CPC G01N 33/6893 (2013.01) [A61K 39/3955 (2013.01); A61K 49/0004 (2013.01); C07K 16/18 (2013.01); G01N 21/66 (2013.01); G01N 21/76 (2013.01); G01N 33/54306 (2013.01); A61K 2039/505 (2013.01); G01N 2458/30 (2013.01); G01N 2800/28 (2013.01); G01N 2800/325 (2013.01); G01N 2800/52 (2013.01)] 26 Claims
OG exemplary drawing
 
1. A method of detecting misfolded transthyretin (TTR) in a biological sample, the method comprising:
(a) contacting a biological sample with a capture antibody that specifically binds to an epitope within residues 89-97 of TTR of SEQ ID NO:26, wherein the capture antibody comprises three Kabat heavy chain CDRs, CDR-H1 of SEQ ID NO:75, CDR-H2 of SEQ ID NO: 76, and CDR-H3 of SEQ ID NO:77, and three Kabat light chain CDRs, CDR-L1 of SEQ ID NO:78, CDR-L2 of SEQ ID NO:79, and CDR-L3 of SEQ ID NO:80, and a reporter antibody that specifically binds to a different epitope of TTR, wherein the reporter antibody is selected from the group consisting of:
an antibody comprising three Kabat/Chothia Composite heavy chain CDRs, CDR-H1 of SEQ ID NO:5, CDR-H2 of SEQ ID NO:7, CDR-H3 of SEQ ID NO:9, and three Kabat/Chothia Composite light chain CDRs, CDR-L1 of SEQ ID NO:11, CDR-L2 of SEQ ID NO:13, and CDR-L3 of SEQ ID NO:15;
an antibody comprising a heavy chain variable region of SEQ ID NO:58 and a light chain variable region of SEQ ID NO:59 or a chimeric version or humanized version thereof;
an antibody comprising three heavy chain CDRs, CDR-H1 of SEQ ID NO: 60, CDR-H2 of SEQ ID NO:61, CDR-H3 of SEQ ID NO:62, and three light chain CDRs, CDR-L1 of SEQ ID NO:63, CDR-L2 of SEQ ID NO:64, and CDR-L3 of SEQ ID NO:65;
an antibody comprising three heavy chain CDRs, CDR-H1 of SEQ ID NO: 66, CDR-H2 of SEQ ID NO:67, CDR-H3 of SEQ ID NO:68, and three light chain CDRs, CDR-L1 of SEQ ID NO:69, CDR-L2 of SEQ ID NO:70, and CDR-L3 of SEQ ID NO:71; and
a polyclonal anti-TTR antibody;
wherein if misfolded TTR is present in the sample, the capture antibody and reporter antibody bind to the misfolded TTR forming a sandwich complex, wherein the biological sample is from a hereditary transthyretin-mediated amyloidosis (ATTR) patient carrying a E89K or E89Q TTR mutation and wherein the misfolded TTR includes the E89K mutation, or the E89Q mutation; and
(b) detecting the reporter antibody that forms a sandwich complex in step (a), if any, to indicate presence or absence of the misfolded TTR.
 
14. A method of detecting misfolded transthyretin (TTR) in a biological sample, the method comprising:
(a) contacting a biological sample with a capture antibody that specifically binds to an epitope within residues 101-109 of TTR of SEQ ID NO:26, wherein the capture antibody comprises three Kabat-Chothia Composite heavy chain CDRs, CDR-H1 of SEQ ID NO:5, CDR-H2 of SEQ ID NO:7, CDR-H3 of SEQ ID NO:9, and three Kabat-Chothia Composite light chain CDRs, CDR-L1 of SEQ ID NO:11, CDR-L2 of SEQ ID NO:13, and CDR-L3 of SEQ ID NO:15, and a reporter antibody that specifically binds to a different epitope of TTR, wherein the reporter antibody is selected from the group consisting of:
an antibody comprising a mature heavy chain variable region of SEQ ID NO: 103 and a mature light chain variable region of SEQ ID NO: 104 or a chimeric version or humanized version thereof,
14G8 (ATCC Deposit No. PTA-124079) or a chimeric version or humanized version thereof,
5A1 ATCC Deposit No. PTA-124080) or a chimeric version or humanized version thereof,
6C1 (ATCC Deposit No. PTA-124077) or a chimeric version or humanized version thereof,
an antibody comprising a heavy chain variable region of SEQ ID NO:58 and a light chain variable region of SEQ ID NO:59 or a chimeric version or humanized version thereof,
an antibody comprising three heavy chain CDR-H1 of SEQ ID NO:60, CDR-H2 of SEQ ID NO:61, CDR-H3 of SEQ ID NO:62, and three light chain CDR-L1 of SEQ ID NO:63, CDR-L2 of SEQ ID NO:64, and CDR-L3 of SEQ ID NO:65,
an antibody comprising three heavy chain CDRs, CDR-H1 of SEQ ID NO: 66, CDR-H2 of SEQ ID NO:67, CDR-H3 of SEQ ID NO:68, and three light chain CDRs, CDR-L1 of SEQ ID NO:69, CDR-L2 of SEQ ID NO:70, and CDR-L3 of SEQ ID NO:71; and
a polyclonal anti-TTR antibody;
wherein if misfolded TTR is present in the sample, the capture antibody and reporter antibody bind to the misfolded TTR forming a sandwich complex; and
(b) detecting the reporter antibody that binds to the misfolded TTR in step (a), if any, to indicate presence or absence of the misfolded TTR.