Large Molecule Bioanalysis for Biologic Drugs, Anti-Drug Antibody (ADA), and Biomarkers

We primarily perform immunological ligand-binding assays (LBA) and enzymatic assays to measure large molecule drugs, ADA, and biomarkers in biological fluids and formulation buffers. Our method validations and sample analyses are compliant with current GLP regulations and FDA guidance on bioanalytical method validation (BMV). While our large molecule bioanalysis services predominantly support GLP preclinical and regulated clinical studies, we also support non-regulated studies, such as exploratory PK and biomarker assessments.

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Bioanalysis for Large Molecules

MicroConstants provides the following services for bioanalysis of macromolecules.

Pharmacokinetic Method Validation and Sample Analysis

  • Most PK sample analyses we conduct use LBA or enzymatic assays that had been validated pursuant to GLP regulations, FDA BMV guidance documents, and current industry standards.
  • We commonly use MSD electrochemiluminescence for LBA because of high sensitivity. ELISA methods are also customary for LBA, while our enzyme methods mostly use fluorescence.
  • We also conduct fit-for-purpose (FFP) “validations,” commonly referred to as a bioanalytical qualification (as distinguished from biomarker qualification for diagnostics).
  • The FFP approach represents a spectrum of assay characterization protocols specifically designed to address the client’s needs, while conserving on expense and time.
  • Either full or FFP validations can be applicable to PK, ADA, and biomarker assays, depending on the needs of the client and the expected level of regulatory review.

Immunogenicity Testing for Anti-Drug Antibody (ADA) and Neutralizing Antibody (NAb), and Characterization of ADA Isotype

  • We can transfer in your ADA assay or make the assay from scratch in our lab. Clients are expected to provide drug reference standard and typically a positive control (PC) antibody. We can subcontract PC antibody production, if necessary, in addition to tending to all other matters required to make an ADA assay for your drug.
  • We evaluate the ADA responses to large and small molecule drugs using a multi-tiered approach as outlined by the 2019 FDA Guidance for Industry: Immunogenicity Testing of Therapeutic Protein Products —Developing and Validating Assays for Anti-Drug Antibody Detection. The multi-tiered approach involves detection of ADA (screening assay), confirmation of ADA specificity by competitive inhibition, and titration of ADA reactivity.
  • Neutralizing ADA (NAb): We have >12 years of experience with NAb, particularly with enzyme therapeutics. Our services can include design and development of a NAb assay in our lab, or transfer-in your method for partial or full validation.
  • Isotypes of ADA: Characterization can include determinations of the predominant ADA isotype(s), such as IgG, IgM, IgA, and IgE; and, if needed, subclasses of IgG.

Biomarker Assays

The most common approach for biomarker analysis is use of commercially available kits. However, we are also experienced with making biomarker assays from “scratch” and with method transfers from clients. We conduct FFP level validations for most biomarker studies and fully GLP compliant validations and sample analyses, when requested by clients for later stage clinical trials. There are two basic formats for biomarker analyses, single analyte (singleplex) and multiplex assays, as described below.

Singleplex Biomarker Assays

  • Single analyte commercial kits are very useful but can vary greatly regarding sensitivity, specificity, consistency, and availability. We have experience with a broad array of biomarkers from many kit and reagent vendors and can identify the best option(s) for your biomarker project.

Multiplex Biomarker Assays

  • Our primary technology for multiplex biomarkers analysis uses MSD electrochemiluminescence to achieve high sensitivity, but we are also capable of doing multiplex fluorescence. We have substantial experience with custom multiplex designs and can expertly help you design a custom biomarker assay to suit your needs.

LC/MS/MS Biomarker Assays

  • Development and validation of bioanalytical methods for the simultaneous quantitation of multiple biomarker peptides or epitope peptides of monoclonal antibodies in serum or plasma matrices.

Pharmacokinetic Analysis & Reports

  • Model-dependent or model-independent PK/TK analysis of bioanalytical data from discovery, preclinical, GLP toxicology, and clinical studies.

Enzymatic Activity Assays

  • Assess the potency of recombinant enzymes/enzyme inhibitors, monitor PD biomarkers, and measure the ADA neutralization activity against recombinant enzymes.
  • Enzyme method validations, like that of LBA PK assay methods, can be fully compliant with current guidance or can be FFP. We model enzyme method validations and sample analyses after current FDA guidance on BMV.

Method Development & Validation for Large Molecule Bioanalysis

We provide full method development and validation for the bioanalysis of large molecules. We primarily perform immunological ligand-binding assays (LBA) and enzymatic assays with method validations and sample analyses that are compliant with current GLP regulations and FDA guidance on bioanalytical method validation (BMV). Our studies predominantly support GLP preclinical and regulated clinical pharmacokinetic (PK), anti-drug antibody (ADA), and biomarker analyses. We also conduct many non-regulated studies, such as for preliminary PK and ADA analyses, as well as exploratory biomarker assessments.

Learn More About Method Development & Validation for Large Molecules

Equipment and Software for Large Molecule Bioanalysis

Our large molecule bioanalytical lab at our San Diego, California headquarters is fully equipped with industry-standard equipment and instrumentation for large molecule bioanalysis. Our scientists have access to the latest incubators, electronic pipettors, plate washers, biosafety cabinets, shakers, centrifuges, refrigerators, freezers, and balances. Our larger instruments include electrochemiluminescence, spectrophotometric and multi-mode fluorescence microplate readers.

See Our Complete Equipment and Software List

Featured Case Study

A Brain-derived Neurotropic Factor (BDNF) Immunoassay Able to Quantitate Both Increased and Decreased BDNF Levels over the Course of Disease

Learn how we established a surrogate matrix to quantitate subnormal levels of brain-derived neurotropic factor (BDNF) in sera, as a pharmacodynamic marker of drug treatment in clinical studies, using ligand-binding assay (LBA) technology.

Read Case Study

Specimen Collection Kits

MicroConstants assembles and distributes protocol-specific specimen collection kits to simplify and streamline the PK collection process for single and multi-site clinical trials. Kits are tailored to your specific sampling needs for the study and can be shipped worldwide. The specimen samples collected can be returned to the testing laboratory of your choice for large molecule bioanalysis.

Learn More About Specimen Collection Kits
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