A vote for robustness: Monitoring serum enzyme activity by thin-layer chromatography of dabsylated bradykinin products

•Robust TLC-based assay for monitoring of peptide cleavage by serum proteases.•Lowered concentrations of dabsylated bradykinin fragment 1–5 are associated with reduced ACE-activity.•Very little patient material (3μL serum) is required.•Simple protocol, no high-end equipment. High-end analytical meth...

Full description

Saved in:
Bibliographic Details
Published in:Journal of pharmaceutical and biomedical analysis 2017-09, Vol.143, p.199-203
Main Authors: Bayer, Malte, König, Simone
Format: Article
Language:English
Subjects:
Angiotensin-converting enzyme
Angiotensin-Converting Enzyme Inhibitors
Bradykinin
Bradykinin - blood
Chromatography, Thin Layer
CRPS
Dabsyl chloride
Humans
Thin-layer chromatography
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:•Robust TLC-based assay for monitoring of peptide cleavage by serum proteases.•Lowered concentrations of dabsylated bradykinin fragment 1–5 are associated with reduced ACE-activity.•Very little patient material (3μL serum) is required.•Simple protocol, no high-end equipment. High-end analytical methods provide excellent data but may lack the robustness required in large analytical studies. In particular complex chemical matrices may cause difficulties and increase the need for extensive sample preparation. For screening of patients we thus developed a low-tech assay to monitor bradykinin degradation by serum proteases. The bradykinin concentration mirrors the activity of angiotensin-converting enzyme (ACE). Dabsylated bradykinin (DBK) and its labeled fragments DBK1-8 and DBK1-5 were visualized by thin-layer chromatography using only 3μL of serum. Lower DBK1-5 levels indicated reduced ACE activity due to medication (ACE-inhibitors) or disease. Provided that purified DBK is available, the assay protocol itself is very simple and does not require any expensive high-end equipment.
ISSN:0731-7085
1873-264X
DOI:10.1016/j.jpba.2017.06.007