PK Assay Development for Biologics Using ELISA/MSD Immunoassay Platforms

Pharmacokinetic study has always been an integral part of the research and development for all the novel therapeutic drugs till date. These pk assay are involved in the investigation of the mechanisms of the course of drug action, its absorption, biodistribution within the human body, metabolism, and finally its excretion. In fact, the pharmacokinetics helps in the understanding both the effectivity as well as safety of the most appropriate dosage form. This dosage form is later subjected for pre-clinical and clinical trials before making it available for mass consumption.

A wide variety of pk bioanalysis studies are employed in characterising the pk properties of biologics both for determining its pre-clinical and clinical applicability. Multiplexing bioanalysis – a single technical process possesses immense ability for generating multiples of several quantitative measurements from a single sample under analysis. Such procedures not only offers good technical efficiency but also reduces the overall time and resource utilization for providing a range of volume limited sample.

Likewise, ELISA has been one of the most promising pk testing methods for ligand binding assay analysis.

Method of ELISA estimation –

The ELISA method has been employed in the detection as well quantification of several biological and chemical molecules such as proteins, peptides, hormones, antibodies, etc. This testing is usually accomplished in a 96 well polystyrene plate. Through ELISA, detection of the molecules proceeds towards completion through the assessment of the conjugated enzyme activity through incubating the desired enzyme with a substrate. This final outcome of this incubation is that we get a measurable signal.

This binding and immobilization ability of the reagents in ELISA makes it relatively easier for separating the bound materials from the non-bound ones throughout the assay duration. Further, it helps in the easy wash-out of the non-specifically bound materials and aids in the measurement of the specific analytes.

ELISA can be accomplished in a variety of mechanisms such as direct ELISA, indirect ELISA, sandwich ELISA, competitive ELISA, and ELISPOT ELISA. Each of this variant has got its own optimal application. Furthermore, ELISA assay can be leveraged across a multitude of research areas such as cardiovascular, oncology, gene therapy, cell therapy, neuroscience, etc. Specifically, this method can be employed in the evaluation of antibodies to specific HLA haplotype thereby supporting the assessment of the both humoral and cellular responses, etc.

Although ELISA is most commonly employed for pk analysis, it has got its own series of limitations as well. PK analysis with mass spectrometry offers specificity, fast development of the analysis method and its validation. Furthermore, the amount of sample required for bioanalysis is relatively less in comparison to the ELISA technique. As an add-on advantage, the technique of mass spectrometry offers characterization of post-translational modification and the ease in assay transfer from animal model study to the analysis of the human sample.

The Meso Scale Discovery or MSD assays can also be employed widely for performing Pharmacokinetic analysis as it yields high throughput, faster results, and is considered to be a cost-efficient method. It is often preferred over its traditional ELISA counterparts due to lower detection limits, and broader dynamic ranges.

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