Method for determining anti-drug antibodies in a minipig sample
Inventors
BROCKHAUS, Janis • Reiser, Astrid • Ritter, Mirko • Stubenrauch, Kay-Gunnar • Vogel, Rudolf • Zadak, Markus
Assignees
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Publication Number
US-11313864-B2
Publication Date
2022-04-26
Expiration Date
Abstract
The present invention relates to in vitro methods for determining the presence or absence of an anti-drug antibody (ADA) or drug/ADA immunecomplexes in a minipig sample. The invention further relates to antibodies specifically binding to minipig IgG that are applicable in the methods of the invention.
Core Innovation
The invention provides an in vitro method for determining the presence or absence of an anti-drug antibody (ADA) in a minipig sample by forming drug/ADA complexes with an excess amount of a drug to which the ADA binds. The formed drug/ADA complexes are contacted with an antibody specifically binding minipig IgG, and the binding of the antibody is determined to decide whether ADA is present in the sample.
The minipig IgG-binding antibody is defined by specific heavy-chain and light-chain CDR1, CDR2, and CDR3 sequences identified by SEQ ID NOs, including CDR set combinations using SEQ ID NO: 3, 4, and 5 with SEQ ID NO: 6, 7, and 8; alternative specified CDR sets using SEQ ID NO: 19, 20, and 21 with SEQ ID NO: 22, 23, and 24; or SEQ ID NO: 27, 28, and 29 with SEQ ID NO: 30, 31, and 32.
The antibody specifically binding to minipig IgG does not bind to human IgG, and in certain embodiments does not bind to mouse IgG. The invention also provides an in vitro method for determining the presence or absence of immune complexes comprising a drug and an antibody specifically binding to the drug by immobilizing the complexes using an antibody specifically binding to the drug and then contacting with the specified minipig IgG-binding antibody.
Claims Coverage
The document includes three independent claims covering three inventive features: an in vitro ADA detection workflow using excess drug to form drug/ADA complexes and probe them with a minipig IgG-specific antibody having defined CDR sequences and no binding to human IgG, an in vitro immune-complex detection workflow using immobilized anti-drug capture and the same minipig IgG-specific antibody, and an antibody specifically binding to minipig IgG defined by alternative heavy/light CDR sets.
Excess-drug drug/ADA complexes contacted with minipig IgG-specific antibody that does not bind human IgG
An in vitro method for determining the presence or absence of an anti-drug antibody (ADA) in a minipig sample by contacting the sample with an excess amount of a drug to form drug/ADA complexes, contacting the complexes with an antibody specifically binding minipig IgG selected from antibodies comprising specified heavy-chain and light-chain CDR1, CDR2, and CDR3 defined by SEQ ID NOs, and determining the binding of the antibody to decide whether ADA is present; the antibody specifically binding to minipig IgG does not bind to human IgG.
Immobilized anti-drug capture of drug/ADA complexes probed with minipig IgG-specific antibody that does not bind human IgG
An in vitro method for determining the presence or absence of immune complexes comprising a drug and an antibody specifically binding to the drug in a minipig sample by immobilizing the drug/ADA complexes with an antibody specifically binding to the drug, contacting the complexes with an antibody specifically binding minipig IgG selected from antibodies comprising specified heavy-chain and light-chain CDR1, CDR2, and CDR3 defined by SEQ ID NOs, and determining binding; the antibody specifically binding to minipig IgG does not bind to human IgG.
Minipig IgG-binding antibody defined by alternative heavy/light CDR sets
An antibody specifically binding to minipig IgG comprising a CDR1 of the heavy chain of SEQ ID NO: 3 with CDR2 of the heavy chain of SEQ ID NO: 4 and CDR3 of the heavy chain of SEQ ID NO: 5, together with a CDR1 of the light chain of SEQ ID NO: 6 with CDR2 of the light chain of SEQ ID NO: 7 and CDR3 of the light chain of SEQ ID NO: 8; or alternative specified heavy-chain and light-chain CDR sets using SEQ ID NO: 19, 20, and 21 with SEQ ID NO: 22, 23, and 24; or SEQ ID NO: 27, 28, and 29 with SEQ ID NO: 30, 31, and 32.
Across the independent claims, coverage centers on forming drug/ADA complexes using excess drug, detecting ADA or immune complexes by probing with a minipig IgG-specific antibody whose heavy- and light-chain CDRs are defined by selected SEQ ID NO combinations, and ensuring the minipig IgG-binding antibody does not bind human IgG. A separate independent claim protects the minipig IgG-binding antibody itself defined by the alternative CDR sets.
Stated Advantages
Low signal-to-noise.
Reliable detection at high drug doses.
Low cross-reactivity.
Documented Applications
In vitro detection of anti-drug antibody (ADA) presence or absence in a minipig sample using drug/ADA complexes and a minipig IgG-specific antibody that does not bind human IgG.
In vitro detection of immune complexes comprising a drug and an antibody specifically binding to the drug in a minipig sample by immobilizing the complexes and probing with the minipig IgG-specific antibody.
Use with minipig sample types including plasma and ophthalmic samples including vitreous humor and aqueous humor.
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