The use of LCMS detection for the bioanalysis of Large Molecules (LM) offers many advantages, including high specificity, wide linear dynamic range, fast method development, and the ability to quantify multiple proteins simultaneously. To fully support the bioanalysis of LM involving hybrid LBA/LCMS, Algorithme Pharma has developed an SOP in line with current industry standards.
Given the potential increase in variability associated with the complexity of LM bioanalytical assays involving highly selective affinity capture enrichment, either prior to, and/or after enzymatic digestion (i.e. hybrid LBA/LCMS approaches), such LM assays require unique validation and sample analysis considerations. Noticeably, they involve the application of acceptance criteria wider than that of small molecules. Algorithme Pharma's SOP outlines the specifics of the validation process of such LM bioanalytical assays in a regulated environment. The main difference between the bioanalysis of small molecules by LCMS and large molecules by hybrid LBA/LCMS is the general acceptance criteria being applied: