Immunogenicity

Screen, confirm, and isotype anti-drug antibodies in a single assay.

Human Multi Tier Anti-Drug Antibody Assay Kit

Product Number: 85096

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Description

The Genalyte Maverick Multi-Tier Anti-Drug Antibody assay (MT-ADA) combines aspects of the traditional tiered immunogenicity testing approach into a single assay.  The MT-ADA assay screens for the presence of positive samples, confirms positive results as ADA, and identifies the isotype (IgM, IgA, IgE, IgG1, IgG2, IgG3, IgG4) at the same time.  Additionally, an anti-rabbit IgG sensor is included on the chip to allow positive control ADAs generated in preclinical species to be used during assay development, and a streptavidin coated sensor is included on the chip as an internal reagent control.

Principle of the Method

Silicon photonic biosensors on the chip surface are functionalized with anti-human isotype capture antibodies to bind ADA by their Fc. As an affinity-purified ADA sample is flowed across the chip, ADAs bind according to their isotype. Then biotinylated drug is flowed over the chip, and binding to true ADAs occurs.  In the final step, streptavidin coated beads bind to bound biotinylated drug amplifying only the true ADA signal.

The MT-ADA multiplex isotype capture assay format directly addresses the weaknesses in other ADA assay formats.

  • The format prevents bridging of soluble target and a false positive result.
  • The format detects monovalent IgG4 ADA, potentially reducing false negative results.
  • The process complies with FDA guidance by detecting and differentiating between all ADA isotypes and sub-classes
  • The assay streamlines the testing workflow by detecting, confirming, and isotypeing ADAs in a single run.
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Background Information

Bridging ELISAs commonly used for Anti-Drug Antibody (ADA) detection requires a bivalent ADA to link either immobilized drug to a labeled drug molecule or two deferentially labeled drug molecules in order to be detected. This format has multiple known disadvantages:

  • It is unable to detect monovalent antibodies (IgG4) since a bridge cannot be formed.
  • It is prone to drug interference.
  • It leads to false positives results due to soluble or dimerized target.

In 2011, The Journal of Immunological Methods reported results of a study titled, “Differential Effect of Drug Interference in Immunogenicity Assays.” In the study, 216 samples from rheumatoid arthritis patients treated with adalimumab were tested for anti-adalimumab antibodies. Bridging ELISAs were not able to detect IgG4 ADAs when compared to a specific IgG4 ADA test (ABT, an antibody binding test) leading to a significant difference in the number of positive samples detected.

  • 7% of samples were positive for ADA by bridging ELISA.1
  • 22% of samples were positive for ADA by the IgG4-specific ABT.1

This type of data demonstrates the importance of detecting all isotypes and supports the use of assay formats which are able to detect IgG4 along with all other ADA isotypes and IgG subtypes.

CITATIONS

1. Hart MH, et al., Differential Effect of Drug Interference in Immunogenicity Assays, Journal of Immunological Methods 372 2011, 196-203.

White Papers

Detection of Isotype of Anti-Adalimumab Antibodies using a Multiplex Photonic Ring Immunoassay
The present study shows the performance characteristics of a human anti-Adalimumab Multi-Tier Anti-Drug Antibody (MT-ADA) assay. This is a multiplex assay that determines the isotype and IgG subclass of ADA in a single run, including IgG4. The sensitivity and free drug tolerance exceed FDA guidelines.

Detection of Isotype of Anti-Infliximab Antibodies using a Multiplex Photonic Ring Immunoassay
The present study shows the performance characteristics of a human anti-Infliximab Multi-Tier Anti-Drug Antibody (MT-ADA) assay. This is a multiplex assay that determines the isotype and IgG subclass of ADA in a single run, including IgG4. The sensitivity and free drug tolerance exceed FDA guidelines.

Detection of Anti-Drug Antibodies in Rabbits with a Multiplex Photonic Ring Immunoassay
The present study shows the performance characteristics of a Rabbit Multi-Tier Anti-Drug Antibody (rMT-ADA) assay. This is a multiplex assay that measures the isotype of anti-drug antibodies (ADA) in a single run. The sensitivity and free drug tolerance exceed FDA guidelines.


Brochures

Detection of all ADA subclass and isotypes, including monovalent IgG4, on a Multiplex Photonic Ring Immunoassay

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