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Protamine-sensitive polymer membrane electrode: characterization and bioanalytical applications.
Anal Biochem 1995; 224(1):212-20AB

Abstract

A polymeric membrane electrode that exhibits significant and analytically useful potentiometric response to submicromolar levels of the heparin antagonist, protamine, is reported. The sensor is prepared by incorporating a lipophilic cation exchanger, potassium tetrakis(4-chlorophenyl)borate (KTpClPB) (at 1 wt%), within a specially formulated polymer membrane composed of 33 wt% 2-nitrophenyl octyl ether (2-NPOE), and 66 wt% poly(vinyl chloride) (PVC). When the polymer film is mounted in an appropriate electrode body, the resulting membrane electrode responds reproducibly to protamine via a nonequilibrium quasi-steady-state change in the phase boundary potential at the membrane/sample interface. Such response can be used to directly monitor, via classical potentiometric titrations, the binding between protamine and a variety of native (porcine and beef) as well as low-molecular-weight heparins. Scatchard analysis of the EMF titration data provides binding constants and stoichiometries for protamine-heparin interactions. The electrode can be further used to follow the enzymatic digestion of protamine by trypsin. In the presence of a given level of protamine, initial rates of potential decrease (-dE/dt) are shown to be linearly related to trypsin activity in solution over the range of 0-130 units/ml. The speed and simplicity of the protamine sensor make it an attractive alternative to classical methods for studying the interaction of protamine with other biologically important macromolecules as well as the proteolytic activity and reaction kinetics of trypsin.

Authors+Show Affiliations

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

Pub Type(s)

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

Language

eng

PubMed ID

7710074

Citation

Yun, J H., et al. "Protamine-sensitive Polymer Membrane Electrode: Characterization and Bioanalytical Applications." Analytical Biochemistry, vol. 224, no. 1, 1995, pp. 212-20.
Yun JH, Meyerhoff ME, Yang VC. Protamine-sensitive polymer membrane electrode: characterization and bioanalytical applications. Anal Biochem. 1995;224(1):212-20.
Yun, J. H., Meyerhoff, M. E., & Yang, V. C. (1995). Protamine-sensitive polymer membrane electrode: characterization and bioanalytical applications. Analytical Biochemistry, 224(1), pp. 212-20.
Yun JH, Meyerhoff ME, Yang VC. Protamine-sensitive Polymer Membrane Electrode: Characterization and Bioanalytical Applications. Anal Biochem. 1995 Jan 1;224(1):212-20. PubMed PMID: 7710074.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Protamine-sensitive polymer membrane electrode: characterization and bioanalytical applications. AU - Yun,J H, AU - Meyerhoff,M E, AU - Yang,V C, PY - 1995/1/1/pubmed PY - 1995/1/1/medline PY - 1995/1/1/entrez SP - 212 EP - 20 JF - Analytical biochemistry JO - Anal. Biochem. VL - 224 IS - 1 N2 - A polymeric membrane electrode that exhibits significant and analytically useful potentiometric response to submicromolar levels of the heparin antagonist, protamine, is reported. The sensor is prepared by incorporating a lipophilic cation exchanger, potassium tetrakis(4-chlorophenyl)borate (KTpClPB) (at 1 wt%), within a specially formulated polymer membrane composed of 33 wt% 2-nitrophenyl octyl ether (2-NPOE), and 66 wt% poly(vinyl chloride) (PVC). When the polymer film is mounted in an appropriate electrode body, the resulting membrane electrode responds reproducibly to protamine via a nonequilibrium quasi-steady-state change in the phase boundary potential at the membrane/sample interface. Such response can be used to directly monitor, via classical potentiometric titrations, the binding between protamine and a variety of native (porcine and beef) as well as low-molecular-weight heparins. Scatchard analysis of the EMF titration data provides binding constants and stoichiometries for protamine-heparin interactions. The electrode can be further used to follow the enzymatic digestion of protamine by trypsin. In the presence of a given level of protamine, initial rates of potential decrease (-dE/dt) are shown to be linearly related to trypsin activity in solution over the range of 0-130 units/ml. The speed and simplicity of the protamine sensor make it an attractive alternative to classical methods for studying the interaction of protamine with other biologically important macromolecules as well as the proteolytic activity and reaction kinetics of trypsin. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/7710074/Protamine_sensitive_polymer_membrane_electrode:_characterization_and_bioanalytical_applications_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(85)71032-9 DB - PRIME DP - Unbound Medicine ER -