Novel Drug Modalities Bioanalysis

Common bioanalytical platforms for macromolecules include:

Liquid Chromatography-Mass Spectrometry (LC-MS) platform: While traditionally used for small molecules, advancements have enabled its use for macromolecules such as oligonucleotides, peptides, and proteins. While triple-quad instruments are used for the quantification of oligonucleotides, peptides, and proteins after enzymatic digestion, high-resolution mass spectrometry (HRMS) can be used for both quantification and characterization of potential biotransformation for macromolecules.

Immunoassays: Immunoassays, or ligand binding assays (LBA), include enzyme-linked immunosorbent assays (ELISAs), chemiluminescent immunoassays (CLIAs, such as MSD), and flow cytometry, etc. These methods analyze an analyte of interest with reagents that specifically bind to the analyte. The analyte is often detected using reagents labeled with enzyme, fluorophore, chromophore, enzyme, fluorophore, chromophore, etc.

RT-qPCR: Real-time quantitative Polymerase Chain Reaction (RT-qPCR) is a laboratory technique used to amplify and simultaneously quantify a targeted DNA, mRNA, oligonucleotide, or other related molecules. It provides a rapid and sensitive method for detecting and quantifying gene expression levels, often used in diagnostic and research settings.

For LBAs, the following elements should be evaluated unless otherwise justified: specificity, selectivity, calibration curve (response function), range (LLOQ to ULOQ), accuracy and precision, dilution linearity, hook effect, and stability. If necessary, carryover or parallelism tests can be conducted.