title: A sensitive and high-throughput LC-MS/MS method for the quantification of pegylated-interferon-alpha2a in human serum using monolithic C18 solid phase extraction for enrichment.
abstract: The analysis of pegylated-interferon-alpha(2a) in patient serum samples is of high interest for clinic research trials, as this therapeutic protein has become an important antiviral treatment. In this study, an LC-MS/MS method for the absolute quantification of pegylated-interferon-alpha(2a) in human serum was developed. The assay achieved a lower limit of quantification of 3.6 ng/mL (60 pM) with the use of a monolithic C(18) solid phase extraction to enrich the target protein. The linear range of the assay was defined up to 54 ng/mL to measure the typical clinical pegylated-interferon-alpha(2a) levels, and within this range, the precision and accuracy were found to be within +/-20%. The method was applied to a clinical study and found suitable for high-throughput analysis of pegylated-interferon-alpha(2a) in human serum. In addition, further investigations suggested the enrichment step may have general application to the sensitive analysis of other low molecular weight proteins.
authors.affiliation: Department of Drug Metabolism and Pharmacokinetics, Novartis Institutes for BioMedical Research, East Hanover, NJ 07936, USA. Ziping.yang@novartis.com
authors.forename: Ziping
authors.initials: Z
authors.lastname: Yang
authors.forename: June
authors.initials: J
authors.lastname: Ke
authors.forename: Michael
authors.initials: M
authors.lastname: Hayes
authors.forename: Matthew
authors.initials: M
authors.lastname: Bryant
authors.forename: Francis L S
authors.initials: FL
authors.lastname: Tse
mesh_terms: D000998:Antiviral Agents / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000097:blood*; D013058:Mass Spectrometry / Q000379:methods*; D011092:Polyethylene Glycols; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity
chemical_list: D000998:Antiviral Agents; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; C100416:peginterferon alfa-2a