Electrochemical assay of protease activities based on polycation/polyanion complex as substrate and polyion sensitive membrane electrode detection
Yükleniyor...
Tarih
2002
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
A novel electrochemical method to detect protease activities is demonstrated. The assay is based on the use of a macromolecular polycation/polyanion substrate; specifically, a complex of the arginine-rich peptide protamine and pentosan polysulfate (PPS), a highly sulfated polysaccharide. As the protease of interest cleaves the protamine within the complex into smaller fragments, free PPS is generated and detected potentiometrically via a polyanion sensitive membrane electrode. Thus, the rate of free PPS generation is proportional to the activity of the protease in the assay solution. The effect of the substrate concentration is examined, as is the influence of the protamine/PPS stoichiometry on the assay performance. Using the optimized composition and concentration of the complex, the determination of trypsin at levels down to 5 U/ml and plasmin at levels approaching 0.002 U/ml can be achieved in a 10 min period. The prospects of further adapting this scheme to determine clot-busting plasminogen activators (e.g. streptokinase, tissue plasminogen activator, etc.) in samples as complex in whole blood are discussed. © 2002 Elsevier Science B.V. All rights reserved.
Açıklama
Anahtar Kelimeler
Plasmin, Plasminogen activators, Polyion sensing, Polymer membrane electrodes, Protease assays, Trypsin
Kaynak
Biosensors and Bioelectronics
WoS Q Değeri
Scopus Q Değeri
Q1
Cilt
18
Sayı
2-3
