期刊
BIOANALYSIS
卷 9, 期 19, 页码 1477-1491出版社
FUTURE SCI LTD
DOI: 10.4155/bio-2017-0105
关键词
C1-INH-HAE biomarker; cleaved high-molecular-weight kininogen; ion-pair 2D-LC-MS/MS; protein biomarker
资金
- Shire
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.
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