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Electrochemical assay of plasminogen activators in plasma using polyion-sensitive membrane electrode detection.
Anal Biochem 1999; 276(1):8-12AB

Abstract

Two relatively simple electrochemical assay methods suitable for the measurement of plasminogen activators (including urokinase (u-PA), streptokinase (SK), and tissue plasminogen activator (t-PA)) in plasma samples are described. In one approach, the initial rate of decrease in the potentiometric response of a polycation-sensitive membrane electrode toward protamine is monitored after addition of a preincubated reaction mixture containing the sample and exogenous plasminogen. The plasmin formed from plasminogen by the activators catalyzes the decomposition of the arginine-rich protamine substrate, yielding smaller polycationic fragments that are not sensed by the electrode. Alternately, the sample, plasminogen, and protamine can be incubated together, and the remaining protamine in this reaction mixture can be measured at a fixed point in time by placing the electrode into the mixture and recording the electromotive force response. Working curves found with both methods for plasma samples spiked with varying levels of the activators cover the expected therapeutic activity ranges found in the plasma of patients treated with these "clot-busting" drugs.

Authors+Show Affiliations

College of Pharmacy, University of Michigan, Ann Arbor, Michigan, 48109-1065, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10585738

Citation

Chang, L C., et al. "Electrochemical Assay of Plasminogen Activators in Plasma Using Polyion-sensitive Membrane Electrode Detection." Analytical Biochemistry, vol. 276, no. 1, 1999, pp. 8-12.
Chang LC, Meyerhoff ME, Yang VC. Electrochemical assay of plasminogen activators in plasma using polyion-sensitive membrane electrode detection. Anal Biochem. 1999;276(1):8-12.
Chang, L. C., Meyerhoff, M. E., & Yang, V. C. (1999). Electrochemical assay of plasminogen activators in plasma using polyion-sensitive membrane electrode detection. Analytical Biochemistry, 276(1), pp. 8-12.
Chang LC, Meyerhoff ME, Yang VC. Electrochemical Assay of Plasminogen Activators in Plasma Using Polyion-sensitive Membrane Electrode Detection. Anal Biochem. 1999 Dec 1;276(1):8-12. PubMed PMID: 10585738.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Electrochemical assay of plasminogen activators in plasma using polyion-sensitive membrane electrode detection. AU - Chang,L C, AU - Meyerhoff,M E, AU - Yang,V C, PY - 1999/12/10/pubmed PY - 2000/2/19/medline PY - 1999/12/10/entrez SP - 8 EP - 12 JF - Analytical biochemistry JO - Anal. Biochem. VL - 276 IS - 1 N2 - Two relatively simple electrochemical assay methods suitable for the measurement of plasminogen activators (including urokinase (u-PA), streptokinase (SK), and tissue plasminogen activator (t-PA)) in plasma samples are described. In one approach, the initial rate of decrease in the potentiometric response of a polycation-sensitive membrane electrode toward protamine is monitored after addition of a preincubated reaction mixture containing the sample and exogenous plasminogen. The plasmin formed from plasminogen by the activators catalyzes the decomposition of the arginine-rich protamine substrate, yielding smaller polycationic fragments that are not sensed by the electrode. Alternately, the sample, plasminogen, and protamine can be incubated together, and the remaining protamine in this reaction mixture can be measured at a fixed point in time by placing the electrode into the mixture and recording the electromotive force response. Working curves found with both methods for plasma samples spiked with varying levels of the activators cover the expected therapeutic activity ranges found in the plasma of patients treated with these "clot-busting" drugs. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/10585738/Electrochemical_assay_of_plasminogen_activators_in_plasma_using_polyion_sensitive_membrane_electrode_detection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(99)94299-9 DB - PRIME DP - Unbound Medicine ER -