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Target interference suppressed anti-drug antibody assay

An anti-drug antibody, antibody technology, applied in biological testing, measuring devices, biological material analysis, etc., can solve problems such as the inability to analyze human samples

Pending Publication Date: 2020-07-14
F HOFFMANN LA ROCHE & CO AG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Here, a specific anti-cynomolgus IgG that does not cross-bind to human IgG was used, thus this assay cannot be used to analyze human samples

Method used

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  • Target interference suppressed anti-drug antibody assay
  • Target interference suppressed anti-drug antibody assay
  • Target interference suppressed anti-drug antibody assay

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Experimental program
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Effect test

Embodiment approach

[0109] Drug interference in ADA assays is a well-known phenomenon, but interference with the target of a drug in an ADA assay is not.

[0110] Typically, after acid dissociation, a neutralization step is followed by acid or base dissociation to allow rebridging of the binding partners, causing interfering factors (if any) in solution to also recombine, which remains a problem.

[0111] A number of protocols have been used to alleviate this problem, such as acid or base dissociation, competitive inhibition with interfering effects of specific antibodies, removal of interfering factors, solid phase extraction with acid dissociation (SPEAD), affinity capture elution ( ACE) and many others. The use of acid dissociation in bridge assays has been shown to somewhat improve drug tolerance to ADA detection (see, e.g., Moxness, M. et al., Clin. Chem. 51(10), 1983; Patton, A et al., J. Immunol. Methods 304 (2005) 189).

[0112] PEG precipitation of target molecules or immune complexes ...

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Abstract

Herein is reported an immunoassay for quantifying the amount of anti-drug antibody, which anti-drug antibody can specifically bind to a drug antibody, which drug antibody can specifically bind to a therapeutic target, in a serum or plasma sample comprising the steps of a) incubating the serum or plasma sample at a pH value that is about the pI value of the target, and optionally removing formed precipitate after the incubation, b) incubating the serum or plasma sample obtained in step a) at a pH value of about 2, and optionally centrifuging the incubated sample to remove formed precipitate, c)adjusting the pH value to about 7.4, adding capture antibody conjugated to a first member of a binding pair and tracer antibody conjugated to a detectable label to the serum or plasma sample obtainedin step b) and incubating the mixture to form a capture antibody-anti-drug antibody-tracer antibody-complex, d) quantifying the complex formed in step c) and thereby quantifying the amount of anti-drug antibody in the serum or plasma sample.

Description

[0001] The present invention belongs to the field of anti-drug antibody assay. Herein is reported an anti-drug antibody assay with reduced interference from the target of a therapeutic drug. Background technique [0002] Moxness, M. et al. (Ann. N.Y. Acad. Sci. USA 1005 (2003) 265-268) reported a radioligand binding assay for total Ig-like insulin antibodies (IAB). Test and control sera were first acidified to dissociate conjugated insulin, and charcoal was added to adsorb serum insulin. After neutralization, the charcoal containing bound insulin was removed from the serum by centrifugation. For each assay, insulin-extracted serum samples were incubated with radiolabeled insulin in the presence and absence of high levels of unlabeled insulin to determine nonspecific and total binding, respectively. Thus, Moxness et al. reported a comparison of two ADA assay protocols in which overnight incubation and acid dissociation were compared. [0003] Patton, A. et al. (J. Immunol. M...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6854G01N33/54393G01N2400/10G01N33/53
Inventor U·达尔G·乔丹R·史塔克M·莫赫森-扎德
Owner F HOFFMANN LA ROCHE & CO AG
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