Application of Automated Liquid Handling in Ligand-Binding Assay-Based Bioanalytical Method Development: the Practical Considerations

Abstract

The ligand-binding assay (LBA) is the standard bioanalytical methodology to support preclinical and clinical development of biologics. LBA methods are based on molecular interactions of specific and selective critical reagents, which are large proteins themselves, with intended analyte in often a complex biological matrix such as serum. There are several benefits of automation in LBA method development. Primarily the replacement of manual with automated liquid handling will result in improved accuracy and precision of the assay [1], reduced human errors and lowering of the physical stress due to manual steps conducted by bench scientists. The decreased variation in pipetting that will then be achieved leads to the generation of higher quality bioanalytical data having greater reproducibility within studies and therefore more comparability across studies over time. This commentary will highlight our experience in applying automated liquid handling to each of these critical components of LBA method development process in bioanalytical laboratories conducting pharmacokinetic (PK), anti-drug antibody (ADA) and target engagement (TE) method development using LBAs, from start of critical reagent screening/pairing, to reagent addition, to sample/quality control (QC) dilutions, preparation of standard calibrators and QC samples with a focus on implementation in regulated bioanalysis. Selection of automation platformsThere is a broad variety of automated liquid handling systems available in the market today. They mainly are either air-based system or liquid-filled system for pipetting with either/or a combination of fixed and disposable tips. Automation platforms with liquid detection system are used to not only provide a record of action of aspiration but also to aid when the volumes of bioanalytical samples are low.Numerous publications have promoted using automated robotic systems in bioanalytical laboratories with different platforms and logistics of set-up [2][3][4][5][6][7]. The 'total laboratory automation (TLA)' proposed for routine clinical chemistry laboratories [4] is not always the best strategy for the application of automation to the development of PK, ADA and TE methods based on LBA methods. Most of the leading manufacturers of integrated robotic and automated liquid handling systems have produced LBA-based systems and have the flexibility to work with multiple assay platforms, run multiple methods in a single operation and are designed to take advantage of the vertical space within a laboratory to achieve the capacity of functionality required.In our laboratory, we took the initial path of purchasing modular automated liquid handling systems which can serve several benefits to our bioanalytical operation. Focus upon the accuracy and precision of liquid handling and its laboratory-wide effect upon flexible operation and reproducibility for LBA methods that will be routinely employed over several years was one of the major reasons for choosing modular platforms over TLA. The breakdown of the preparation of ...

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