The Complete Guide to Antibody-drug Conjugates Bioanalysis

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The Complete Guide to Antibody-drug Conjugates
Bioanalysis
www.creative-biolabs.com/adc/
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Content
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ADC Heterogeneity
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DAR Values in vivo Dynamic Changes
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How to Choose Samples
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How to Choose Bioanalysis Platform
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ADC Heterogeneity
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1 ADC Heterogeneity
Antibodies and small molecules in ADCs are
covalently linked through a linker that allows
small molecule drugs to bind to the lysine side
chain amino groups in the antibody, such as
T-DM1, or to sulfhydryl groups obtained by
reduction of disulfide bonds between antibody
chains, such as an anti-CD30 IgGl monoclonal
antibody cAClO-MMAE conjugate or to an engineered
cysteine residue introduced at a particular site
on the antibody, such as a THIOMAB-drug
conjugate. The number of small molecules in the
ADC and the location of the connections can be
inconsistent through the first two forms, and the
ADC is actually a mixture.
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• It has been reported that the drug-to-antibody
  ratio (DAR) of T-DM1 is between 0 and 8 with an
  average DAR value of 3.5 and the ADC DAR values
  obtained by interchain disulfide bond reduction
  are usually even. ADCs obtained by introducing
  engineered cysteines at specific sites on the
  antibody are less heterogeneous. As reported in
  the literature, homologous ADCs with a DAR value
  of 2 can be obtained.

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DAR Values
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2 DAR Values in vivo Dynamic Changes
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Samples
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How to Choose Samples

• For small molecule drugs and protein drugs, the
  relationship between exposure and response
  (exposure - response, ER) can be figured out by
  evaluating prototype drugs. However, the presence
  of both antibodies and small molecule drugs in
  the ADC results in a dynamic change in the DAR
  value in the body. Combined with the current
  limited clinical data of the ADC, it is difficult
  to determine which test substance determines or
  affects the ER relationship of the ADC. In
  general, in the early stages of ADC development,
  as many as possible to determine a variety of
  analytes in the ADC to understand the various
  components of the PK characteristics.

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How to Choose Samples

• The combined information of the samples helps to
  understand the ER relationship of the ADC. In
  addition, DAR conjugation measurements can
  provide information on changes in the ADC that
  help to select the appropriate analytical
  technique. In addition, the introduction of ADC
  as a biological macromolecule into the body may
  cause immunogenicity and produce anti-therapeutic
  antibody (ATA). ATA may lead to accelerated ADC
  clearance or reduced potency of ADC.
  Alternatively, ADC may be taken to other sites by
  ATA. Therefore, it is necessary to evaluate ATA
  during ADC development.

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Bioanalysis Platform
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How to Choose Bioanalysis Platform

• Traditionally, in vivo quantitative analysis of
  biological macromolecules using ELISA method,
  small molecule drugs using LC-MS / MS method
  27. As mentioned above, both the antibody and
  the small molecule drug moiety need to be
  biologically quantified during the development of
  the ADC. Therefore, the methods of ELISA and
  LC-MS / MS have become common methods for ADC
  analysis.

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