# US 2021/0239709 A1 · Improved immunogenicity assays > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0239709 A1 - Document ID: US20210239709A1 - Status: Published application - Kind: A1 - Application number: 16/495,131 - Filed: 2018-03-29 - Published: 2021-08-05 - Earliest claimed: 2018-03-29 - Assignees: Ablynx N.V. - Applicants: Ablynx N.V. - Inventors: Annelies Coddens; Veerle Snoeck; Marie-Ange Buyse; Carlo Boutton - Web page: https://patentgrep.com/patent/US20210239709A1 ## CPC - G01N 33/54333: {Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction} - G01N 33/6857: {Antibody fragments} - C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® - G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention relates to improved immunogenicity assays and methods for performing the same. In particular, the present invention relates to improved immunogenicity assays that are capable of distinguishing neutralizing anti-drug antibodies from non-neutralizing anti-drug antibodies. More in particular, the present invention relates to improved immunogenicity assays that allow (amounts and/or concentrations of) neutralizing anti-drug antibodies to be detected and/or measured in a sample even in the presence of non-neutralizing anti-drug antibodies. ## Claims ### Claim 1 Method for detecting and/or measuring, in a sample, anti-drug antibodies (ADA) against an ISV-based drug, in which said ISV-based drug comprises one or more immunoglobulin single variable domains (ISVDs), in which each such ISVD is comprised of framework regions (FRs) and complementarity determining regions (CDRs) and in which at least one of the ISVDs present in the ISV-based drug is specific for a therapeutically relevant target, said method comprising performing a bridging anti-drug antibody assay in the presence of a variant of said ISV-based drug, which variant is such that it is non-functional with respect to binding to the therapeutic target(s) to which the ISV-based drug is intended to bind (but otherwise is essentially identical to the ISV-based drug). ### Claim 2 (depends on 1) Method according to claim 1, in which the bridging anti-drug antibody assay is performed in the presence of a variant of said ISV-based drug, which variant is such that it is not capable of specifically binding to the therapeutic target(s) to which the ISV-based drug is intended to bind (but otherwise is essentially identical to the ISV-based drug). ### Claim 3 (depends on 1, 2) Method according to claim 1 or 2, in which the bridging anti-drug antibody assay is performed in the presence of a variant of said ISV-based drug, wherein said variant is essentially identical to the ISV-based drug, except that the one or more ISVs in the variant that correspond to the one or more ISVs that, in the ISV-based drug, are intended to bind to the (one or more) therapeutic targets of the ISV-based drug have CDR's that are different from the CDR's in the corresponding ISVs in the ISV-based drug, such that said variant is non-functional with respect to binding to the therapeutic target(s) to which the ISV-based drug is intended to bind (but otherwise is essentially identical to the ISV-based drug). ### Claim 4 (depends on 1, 2, 3) Method according to any of claims 1 to 3, in which the bridging anti-drug antibody assay is performed in the presence of a variant of said ISV-based drug, wherein said variant is essentially identical to the ISV-based drug, except that the one or more ISVs in the variant that correspond to the one or more ISVs that, in the ISV-based drug, are intended to bind to the (one or more) therapeutic targets of the ISV-based drug have CDR's that are different from the CDR's in the corresponding ISVs in the ISV-based drug, such that said variant is not capable of specifically binding (as defined herein) to the therapeutic target(s) to which the ISV-based drug is intended to bind (but otherwise is essentially identical to the ISV-based drug). ### Claim 5 Method according to any of the preceding claims, in which the variant has essentially the same amino acid sequence as the ISV-based drug, except for the changes to the CDRs that, in the variant, remove the ability of the ISV(s) present in the variant to bind to the therapeutic target(s) of the ISV-based drug. ### Claim 6 Method according to any of the preceding claims, in which the variant has a degree of sequence identity (as defined herein) with the ISV-based drug of at least 90%, such as at least 95%, in particular at least 99% (up to and including 100%), not taking into account the changes in the CDRs that, in the variant, remove the ability of the ISV(s) present in the variant to bind to the therapeutic target(s) of the ISV-based drug. ### Claim 7 Method according to any of the preceding claims, in which performing the bridging anti-drug antibody assay comprises the steps of: a) contacting said sample with a capturing agent that is immobilized on a support, wherein said capturing agent is or essentially consists of said ISV-based drug, under conditions such that any anti-drug antibodies against ISV-based drug can bind to said capturing agent; b) (optionally) removing any components or constituents present in said sample that do not bind to the capturing agent; c) detecting or measuring any anti-drug antibodies that have bound to the capturing agent, by contacting the complex of the capturing agent and any captured anti-drug antibodies with a detection agent, which detection agent is or essentially consists of said ISV-based drug which is linked to a detectable label, under conditions such that said detection agent can bind to (the complex of the capturing agent and) any captured anti-drug antibodies. ### Claim 8 (depends on 7) Method according to claim 7, in which in step c), essentially only neutralizing ADAs against the ISV-based drug are detected or measured. ### Claim 9 Method according to any of the preceding claims, which is performed on a sample that contains (or is suspected to contain), both neutralizing and non-neutralizing ADAs against the ISV-based drug. ### Claim 10 Method according to any of the preceding claims, which is performed on a sample obtained from a human subject. ### Claim 11 Method according to any of the preceding claims, which is performed on a sample of whole blood, serum, plasma, lymph fluid, ocular fluid, bronchoalveolar fluid/BALF, cerebrospinal fluid or another biological fluid (such as sputum or nasal washes); and in particular a sample of whole blood, serum or plasma. ### Claim 12 Method according to any of the preceding claims, in which the ISV-based drug has a half-life in man (expressed as t ½ beta) of at least 3 days. ### Claim 13 Method according to any of the preceding claims, in which the ISV-based drug comprises at least two ISVs, such as 2, 3, 4 or 5 ISVs ### Claim 14 (depends on 13) Method according to claim 13, in which the ISV-based drug comprises at least one ISV against serum albumin.