Antibodies for Universal PK Assays in Human IgG Drug Development

Antibodies Inc. is excited to announce the launch of our new antibodies proven for the development of "generic" or "universal" pharmacokinetic (PK) assays to support biologic drug development!

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At Antibodies Inc., we specialize in providing top quality antibodies to scientists worldwide. These critical reagents enable accurate assay development and sample testing, designed to enhance drug safety and efficacy studies in biologic drug development. Our antibodies will provide for rapid and accurate determination of biologic drug concentration in preclinical studies.

These antibodies are designed to streamline assay development and sample testing, providing researchers with the precision they need to enhance drug safety and efficacy studies in biologic drug development. The antibodies are compatible with any immunoassay platform including ELISA, Gyros, Meso Scale Discovery (MSD), AlphaLISA and Ella.

Our antibodies feature:

• Long literature citation history
• Biologic drug (IgG) concentration determination (PK) - mouse, rat, and NHP
• Exploits the minimal dissimilarity in IgG structure between humans and preclinical species
• Useful in early toxicology studies
• Assists in drug candidate selection process
• Minimal assay development or optimization time required
• Rapid results in a non-GLP environment

Introduction to Generic and Universal Pharmacokinetic (PK) Assays

Generic PK assays, also called PK assays, are used in early drug discovery and development to provide support for monoclonal antibody drug candidates (human IgG) and antibody drug conjugates (ADC's) that are dosed in preclinical species. Generic PK assays are immunoassays (often referred to as ligand binding assays, or LBA for short) that measure the concentration of a dosed biologic drug at specific time intervals after dosing. The reason generic assays are used at this point in the early drug discovery process is because lead candidates are in the process of being selected. Once the lead candidates are selected, anti-idiotype antibodies will be developed and used in the building of assays for use in humans. Anti-idiotype antibodies bind to the variable region of another antibody and allows for the detection of human IgG antibody drugs in human matrix (serum and plasma) that contains vast amounts of human IgG.

Key Characteristics of a Generic PK Assay

• Specificity - the generic PK assay should only detect human IgG despite the structural similarities with the IgG of mouse, rat and non-human primates.
• Short method development time - generally only a few days are required for method development.
• Flexible - can be used for human IgG1, IgG4 and antibody drug conjugates of the IgG1 and IgG4 isotypes
• Platform agnostic - antibodies from which generic PK assays are developed and validated should function well on the commonly used immunoassay platforms including ELISA, Meso Scale Discovery (MSD) and Gyros (1).

Table 1. Listing of useful antibodies for generic PK assay development and validation. NHP = non-human primate

Pharmacokinetics

Pharmacokinetics is a branch of pharmacology that studies what happens to a drug once dosed in an animal or a human. Immunoassays are used to determine the concentration of a biologic drug once dosed over time. For example, blood is drawn at specific time intervals (time zero, 1 hour, 4 hours, 1 day, 2 days, 3 days ...), processed to serum or plasma, and the concentration is plotted over time. Drug concentrations are indicative of the level of effectiveness and drug concentration information is a contributing factor in the determination of dosing levels for future studies.

Figure 1. A representative PK profile, plotting drug concentration verses time using a colorimetric ELISA assay. The mean serum concentration times profiles of healthy volunteers following a single subcutaneous injection of 40 milligrams of adalimumab and a biosimilar of adalimumab (2).

Toxicokinetics

In pre-clinical toxicology studies, drugs are dosed at high concentrations to assess potential toxicity. These toxicokinetics (TK) studies are similar to pharmacokinetics (PK) studies with a few key differences. First, TK studies are performed at very high doses, allowing toxic affects to be more apparent. Second, TK studies are only run in animals - not in humans. Additionally, the goal of TK studies is to determine the levels where toxic effect are observed and to guide dosing recommendations for subsequent studies. PK studies determine drug efficacy by determining clearance and bioavailability within the biological system.

Resources in the Literature

There are several resources in the literature that can be very helpful. In 2013 a group at Pfizer in Andover used clone JDC-10 on 4 different immunoassay platforms (ELISA, MSD, Gyrolab and AlphaLISA (1). JDC-10 has been used in a homogeneous no-wash format as the capture and detection simultaneously (3) and as capture and detection antibodies on the Gyros platform (5).

Related Services

• At Antibodies Inc., we take pride in custom generation of the highest quality Positive Control Antibodies for Anti-Drug Antibody (ADA) assay development. From custom antibody generation to purification and characterization, we ensure the success of your assay development projects.

• We offer ADCC/CDC assay services to evaluate antibody and complement activity against target cell lines. Our assays provide dose-dependent kinetic responses, with optimized E/T ratios and activity measurements across antibody concentrations.

Related Products

Coating Buffers: Coating buffers optimize antibody or antigen adsorption onto ELISA plates, stabilize tertiary structures, extend shelf life, and minimize capture reagent usage.

Blocking Buffers: Block buffers target antibody-sandwich or antigen-down ELISAs, preserve hydration of dried coat proteins, mitigate non-specific binding, enhance protein stability, and decrease assay variability for prolonged shelf life.

Sample and Assay Diluents: ICT's Sample Diluents are suitable for antibody-sandwich or antigen-down ELISAs. They minimize non-specific binding interactions during the assay process, reduce sample variation and matrix interference, and decrease background noise.

• Protein-Free Sample Diluent: a protein-free matrix used for the dilution of biological samples(e.g. serum, cell culture media) into the useful range of antibody-sandwich or antigen-down ELISA format assays. This unique buffer contains a heterogeneous mixture of proprietary molecules that help reduce background noise associated with non-specific bridging of signal-generating conjugates to the plate well surface.

• Neptune Assay Diluent: formulated to address non-specific binding issues with human, porcine, or bovine serum or plasma samples. This non-mammalian assay additive works to enhance the specific anti-analyte/antigen antibody signal.

• Assay Diluent Optimization Pack: our optimization pack contains 4 x 100 mL ELISA development buffers (General Assay Diluent, IgM-Reducing Assay Diluent, Neptune Assay Diluent, and Antigen-Down Assay Diluent) and offers an economical method of addressing matrix effects and optimizing signal-to-noise ratio in ELISA development projects.

• Sample Diluent Optimization Pack: our optimization pack provides 4 x 100 mL bottles of Sample Diluent formulations (General Serum Diluent, Plasma Sample Diluent, Sample Diluent - Non Mammalian-Based (formerly Neptune Sample Diluent), and Protein-Free Sample Diluent) for an economical and fast method to screen for the best Sample Diluent for an assay.

Substrates and STOP Solutions: Substrates improve ELISA sensitivity, minimize background noise, ensure reproducible linearity, and are eco-friendly, while STOP solutions standardize readings, halt color development, and reduce variation for enhanced reproducibility.

Coming Soon!

We will soon release a colorimetric ELISA that can be used to measure human IgG drugs dosed in preclinical species.

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References

1. Leary BA, Lawrence-Henderson R, Mallozzi C, Fernandez Ocaña M, Duriga N, O'Hara DM, Kavosi M, Qu Q, Joyce AP. Bioanalytical platform comparison using a generic human IgG PK assay format. J Immunol Methods. 2013 Nov 29;397(1-2):28-36.
2. Hyland E, Mant T, Vlachos P, Attkins N, Ullmann M, Roy S, Wagner V. Comparison of the pharmacokinetics, safety, and immunogenicity of MSB11022, a biosimilar of adalimumab, with Humira(®) in healthy subjects. Br J Clin Pharmacol. 2016 Oct;82(4):983-93.
3. Microsoft Word - generic_pharmacokinetic_assay_SPARCL.docx (lumigen.com)
4. Carrasco-Triguero M, Davis H, Zhu Y, Coleman D, Nazzal D, Vu P, Kaur S. Application of a Plug-and-Play Immunogenicity Assay in Cynomolgus Monkey Serum for ADCs at Early Stages of Drug Development. J Immunol Res. 2016;2016:2618575.
5. Joyce AP, Wang M, Lawrence-Henderson R, Filliettaz C, Leung SS, Xu X, O'Hara DM. One mouse, one pharmacokinetic profile: quantitative whole blood serial sampling for biotherapeutics. Pharm Res. 2014 Jul;31(7):1823-33.