Protamine-responsive polymeric membrane electrode
First Claim
1. An ion-responsive electrode membrane comprising, in admixture, a polymeric matrix material, lipophilic cation-exchange material, and, optionally, a plasticizer, the electrode membrane admixture formulation being selectively responsive to cationic hydrophobic macromolecules.
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Accused Products
Abstract
A polymeric membrane electrode gives potentiometric responses to macromolles, such as protamine, in solution or biological specimens. A protamine-responsive membrane includes a lipophilic cation-exchange material for ionic macromolecules in a specially formulated plasticized, polymeric matrix material. Potentiometric response to protamine has been observed in membranes comprising 30-70 wt. % polymeric matrix material, such as polyvinyl chloride; 1-3 wt. % salt of a tetraphenylborate, such as potassium tetrakis(4-chlorophenyl) borate or salt of an organophosph(on)ate, such as calcium bis-[4-(1,1,3,3-tetramethylbutyl)phenyl] phosphate; and 30-70 wt. % of a plasticizer, such as 2-nitrophenyl octyl ether or tris (2-ethylhexyl) phosphate. The potentiometric response of these membranes to protamine can be used to study the binding interaction of protamine with heparin (both native and low molecular fractions) or to monitor protease activity, for example, by following the rate of degradation of intact protamine or a synthetic substrate by trypsin, as well as to measure protamine concentration in biological fluids, such as blood or plasma.
27 Citations
30 Claims
- 1. An ion-responsive electrode membrane comprising, in admixture, a polymeric matrix material, lipophilic cation-exchange material, and, optionally, a plasticizer, the electrode membrane admixture formulation being selectively responsive to cationic hydrophobic macromolecules.
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16. An ion-responsive electrode membrane prepared by the process comprising:
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(a) forming an ion-responsive membrane comprising, in admixture, a polymeric matrix material and an ion-exchange material, and, optionally, a plasticizer, the ion-responsive membrane admixture formulation being responsive to a macromolecular analyte; (b) casting the ion-responsive membrane into a film; (c) conditioning the film with a concentrated solution of analtye ions for a period of at least 12-15 hours; and (d) exposing the film to a high-ionic strength solution for a short period of time to deplete the surface of the conditioned film of analyte ions.
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17. A cation-responsive electrode membrane comprising, in admixture:
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0.1-5 wt % lipophilic cation-exchange material selective to cationic hydrophobic macromolecules; 30 to 70 wt % plasticizer; and 30 to 70 wt % polymer matrix material, the admixture formulation being optimized to selectively sense cationic hydrophobic macromolecules. - View Dependent Claims (18, 19)
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20. A protamine-responsive polymeric membrane comprising:
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1 wt % potassium tetrakis(4-chlorophenyl) borate; 49.5 wt % 2-nitrophenyl octyl ether; and 49.5 wt % poly(vinyl chloride).
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21. A protamine-responsive polymeric membrane for sensing protamine in biological fluids comprising:
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1.0 wt % bis-[4-(1,1,3,3-tetramethylbutyl)phenyl] phosphate calcium salt; 49.5 wt % tris (2-ethylhexyl) phosphate; and 49.5 wt % PVC.
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22. A membrane electrode having an ion-responsive membrane, the membrane electrode comprising:
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(a) a housing containing a reference solution; (b) an electrode arranged in the housing so that it is disposed in the reference solution; and (b) an ion-responsive membrane of a polymeric matrix material which is non-reactive with the liquid medium, an ion-exchange material, and, optionally, a plasticizer, the ion-responsive membrane being selective to protamine, the ion-responsive membrane being disposed on one end of the housing to seal the reference solution inside the housing and to contact a sample solution external to the housing.
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23. A method of measuring the concentration of protamine in a liquid medium as a function of its potentiometric response, the method comprising:
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(a) bringing a membrane electrode having an ion-responsive membrane into contact with the liquid medium containing an unknown quantity of protamine, the ion-responsive membrane being characterized in that it comprises a polymeric matrix material which is non-reactive with the liquid medium, an ion-exchange material, and a plasticizer; and (b) measuring a potentiometric response which is indicative of the concentration of protamine in the liquid medium. - View Dependent Claims (24, 25, 26)
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27. A method of monitoring protease activity comprising the steps of:
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(a) adding a proteolytic enzyme to a liquid medium containing a known amount of protamine or a synthetic peptide as a substrate for the proteolytic enzyme; and (b) measuring the change in concentration of protamine or synthetic peptide in the liquid medium as a function of its potentiometric response as the proteolytic enzyme degrades protamine into smaller fragments which do not produce a potentiometric response. - View Dependent Claims (28)
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29. A solid state membrane electrode for measuring the concentration of cationic macromolecules in a liquid medium as a function of potentiometric response comprising a conductive substrate and a membrane material arranged to be in adherence with said conductive substrate, said membrane material comprising a polymeric matrix material which is non-reactive with the liquid medium, an ion-exchange material, and, optionally, a plasticizer, said membrane being responsive to the cationic macromolecule.
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30. A method of monitoring protease activity comprising the steps of:
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(a) adding a proteolytic enzyme to a liquid medium containing a known amount of protamine or a synthetic peptide having an amino acid sequence with at least an arginine trimer and an arginine dimer sequence connected by a hydrophobic amino acid residue as a substrate for the proteolytic enzyme; and (b) measuring the change in concentration of protamine or synthetic peptide in the liquid medium as a function of its potentiometric response as the proteolytic enzyme degrades protamine into smaller fragments which do not produce a potentiometric response.
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Specification