Internal standards in the quantitative determination of protein biopharmaceuticals using liquid chromatography coupled to mass spectrometry

摘要: Following the increase in development of protein biopharmaceuticals, there is a growing demand for sensitive and reliable quantification these proteins complex biological matrices such as plasma serum to support (pre)-clinical research. In this field, ligand binding assays (LBAs) are currently standard analytical technique, but recent years, trend towards use liquid chromatography hyphenated with (tandem) mass spectrometry (LC-MS/MS). One reasons possibility internal standards correct variability thus improve precision accuracy results. LC-MS/MS bioanalysis small molecules, standardization quite straightforward: either stable-isotope labeled (SIL) form analyte or structural analogue used. For biopharmaceutical proteins, situation more complex. Since interest digested mixture peptides, one which subsequently used quantification, options standardization. A SIL intact signature peptide can be addition, modified SIL-peptide standard, containing cleavable groups possibility, an generated during analysis by using differential derivatization techniques. paper we provide overview different field absolute targeted biopharmaceuticals LC-MS/MS, based on literature from 2003 2011. The advantages disadvantages approaches evaluated both regard correction they steps their generic availability. As most lead acceptable results terms precision, conclude that no clear preferable method LC-MS/MS. It essential, however, any step not covered chosen, should carefully optimized controlled.