Journal
BIOANALYSIS
Volume 11, Issue 8, Pages 785-795Publisher
FUTURE SCI LTD
DOI: 10.4155/bio-2018-0277
Keywords
critical reagents; endogenous biomarker; hybrid LC-MS assay; immunocapture (IC); ligand-binding assay (LBA); surrogate peptide
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In recent years, hybrid ligand-binding assays (LBAs)/LC-MS assays have been increasingly used for quantitation of protein biomarkers in biological matrices. However, unlike in LBAs where the importance of critical reagent screening and characterization is well understood and widely reported, benefits of well-characterized hybrid LC-MS assay reagents are frequently underestimated. Two groups of analyte-specific reagents, binding reagents and assay calibrators, are considered the critical reagents for biomarker assays. In this article, we summarize the similarities and differences of critical reagents used in LBAs and hybrid LC-MS assays, overview the benefits and approaches of critical reagent screening, characterization, antibody conjugation and discuss bioanalytical considerations in hybrid LC-MS assay development for robust measurements of protein biomarkers in biological matrices.
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