Fluorescence Detection by HPLC-FLD of Antibodies and Other Complex Proteins at Low Concentrations Utilizing Intrinsic Fluorescence for Reliable Protein Characterization
Presented by Sydney Huffman at PharmSci 360, this poster highlights a validated HPLC-fluorescence detection (HPLC-FLD) method for detecting and quantifying low concentrations of antibodies and complex proteins without fluorescent labeling. By leveraging the intrinsic fluorescence of aromatic amino acids, the method offers greater sensitivity than conventional UV detection, enabling more accurate characterization of monomeric species and protein purity at lower sample concentrations. The method demonstrated strong performance across key validation parameters, including repeatability, linearity, accuracy, and specificity, meeting GLP acceptance criteria. Findings support HPLC-FLD as a reproducible, cost-effective alternative to more complex analytical approaches for protein characterization and generating certificates of analysis (CoAs).
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