US 12,480,957 B2
Method of screening for anti-drug antibodies against immunoglobulin single variable domain-based drugs
Annelies Coddens, Oudenaarde (BE); Veerle Snoeck, Zingem (BE); Marie-Ange Buyse, Merelbeke (BE); and Carlo Boutton, Wielsbeke (BE)
Assigned to Ablynx N.V., Ghent-Zwijnaarde (BE)
Appl. No. 16/495,131
Filed by Ablynx N.V., Ghent-Zwijnaarde (BE)
PCT Filed Mar. 29, 2018, PCT No. PCT/EP2018/058232
§ 371(c)(1), (2) Date Sep. 18, 2019,
PCT Pub. No. WO2018/178307, PCT Pub. Date Oct. 4, 2018.
Claims priority of provisional application 62/479,496, filed on Mar. 31, 2017.
Prior Publication US 2021/0239709 A1, Aug. 5, 2021
Prior Publication US 2022/0099682 A9, Mar. 31, 2022
Int. Cl. G01N 33/68 (2006.01)
CPC G01N 33/6857 (2013.01) [C07K 2317/569 (2013.01)] 14 Claims
 
1. A method comprising:
a) selecting an immunoglobulin single variable domain (ISV)-based drug, wherein the ISV-based drug comprises a polypeptide having an amino acid sequence of one or more ISV(s), each ISV comprising four framework regions (FW1, FW2, FW3, and FW4) and three complementarity determining regions (CDR1, CDR2 and CDR3);
b) producing an ISV-based drug variant that comprises a structure that is essentially identical to the ISV-based drug, wherein the ISV-based drug variant has, relative to the ISV-based drug, one or more amino acid changes in a CDR(s);
c) contacting a sample with the ISV-based drug variant and a first agent, wherein said first agent is immobilized on a support and comprises said ISV-based drug, and wherein said contacting occurs under conditions such that any neutralizing ADAs against the ISV-based drug can bind to the ISV-based drug of the first agent;
d) washing the first agent to remove components or constituents present in said sample that are not bound to the first agent; and
e) contacting the first agent and any neutralizing ADAs bound to the ISV-based drug of the first agent with a second agent, wherein the second agent comprises said ISV-based drug linked to a detectable label, wherein said contacting occurs under conditions such that the ISV-based drug of said second agent can be bound by any neutralizing ADA against the ISV-based drug that are bound to the ISV-based drug of the first agent.