2D-LC–MS/MS to measure cleaved high-molecular-weight kininogen in human plasma as a biomarker for C1-INH-HAE. (October 2017)

• Record Type: Journal Article
• Authors: Zhang, Guodong; Sexton, Daniel J; Faucette, Ryan R; Qiu, Yongchang; Wu, Jiang
• Is Part Of: Bioanalysis. Volume 9:Number 19(2017)
• Page Start: 1477
• Page End: 1491
• Publication Date: 2017-10
• Subjects: C1-INH-HAE biomarker -- cleaved high-molecular-weight kininogen -- ion-pair 2D-LC–MS/MS -- protein biomarker; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2017-0105
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: C1-INH-HAE is caused by activation of plasma kallikrein which subsequently cleaves high-molecular-weight kininogen (HMWK) to generate bradykinin and cHMWK.Materials & methods: A novel ion-pair 2D LC–MS/MS assay was developed to measure the 46 kDa cHMWK in plasma as a biomarker for C1-INH-HAE. The sample preparation included sodium dodecyl sulfate denaturation, methanol crash, chymotryptic digestion and peptide enrichment by solid phase extraction.Results: The LLOQ was 200 ng/ml. The overall cHMWK recovery combining crash and digestion was 57.5%. The precision of the method was ≤12.7% and accuracy ≤-13.8%.Conclusion: A reagent-free LC–MS assay has been developed for the quantitation of 46 kDa cHMWK, which was shown to be elevated in plasma of C1-INH-HAE patients due to C1-INH deficiency relative to that of healthy subjects.