Sorbent composition for pre-analytical treatment of samples
AssigneeImmundiagnostik AG
InventorsFranz-Paul Armbruster, Anja Helmschrodt
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Abstract
A method for pre-analytical treatment of a serum or plasma sample from a patient suspected of suffering from oxidative stress, which includes contacting the sample with one or more microcapsules having a gelled alginate core and a semipermeable coating, where the alginate core includes dispersed receptors against an oxidised human parathyroid hormone (PTH) peptide. The semipermeable membrane can be obtained by layer-by-layer deposition of polycationic and polyanionic macromolecules onto the gelled core, following by hardening, crosslinking and co-acervation of the macroionic phases.
Claims10
A method of obtaining a sorbent material for immunological pre-treatment of a biological liquid, wherein the sorbent material comprises microcapsules obtainable by:
- (i) solubilizing antibodies in a physiological buffer containing 0.3 to 2.5 percent by weight alginic acid producing a solution and dropping the solution into an aqueous buffer containing ions of a two-valent salt to precipitate alginate cores with diameters from 0.5 to 5.0 millimeters and volumes ranging from 0.1 to 50 microlitres;
- (ii) alternately immersing the alginate cores in a polycationic macropolymer liquid buffer solutions and a polyanionic macropolymer liquid buffer solution to achieve a layer-by-layer electrostatic deposition and form macroion-rich liquid-liquid phases on the alginate cores;
- (iii) hardening said macroion-rich liquid-liquid phases by complex coacervation at ambient temperatures for a period of time to obtain microcapsules consisting of precipitated alginate cores each surrounded by a multilayered macroion membrane;
- (iv) lyophilizing the microcapsules to yield multilayer macroion membranes that are permeable to proteins with molecular weights no greater than about 100,000 Daltons producing semipermeable microcapsules;
- and
- (v) conditioning said semipermeable microcapsules and, optionally, classifying the semipermeable microcapsules to render them suitable for addition to biological fluids for immunological treatment and removal of peptidic components smaller than 100,000 Daltons.
A method of pretreating plasma or serum in an automated test system for removal of components interfering with a quantitative measurement of analyte, the method comprising:
- contacting said plasma or serum with microcapsules obtained from a method comprising:
- (i) Solubilizing antibodies in a Physiological buffer containing 0.3 to 2.5 Percent by weight alginic acid producing a solution and dropping the solution into an aqueous buffer containing ions of a two-valent salt to precipitate alginate cores with diameters from 0.5 to 5.0 millimeters and volumes ranging from 0.1 to 50 microliters;
- (ii) Alternately immersing the alginate cores in a polycationic macropolymer liquid buffer solution and a polyanionic macropolymer liquid buffer solution to achieve a layer-by-layer electrostatic deposition and form macroion-rich liquid-liquid phases on the alginate cores;
- (iii) Hardening said macroion-rich liquid-liquid phases by complex coacervation at ambient temperatures for a period of time to obtain microcapsules consisting of precipitated alginate cores each surrounded by a multilayered macroion membrane;
- (iv) Lyophilizing the microcapsules to yield multilayer macroion membranes that are permeable to proteins with molecular weights no greater than about 100,000 Daltons, producing semipermeable microcapsules;
- and
- (v) Conditioning said semipermeable microcapsules and, optionally, classifying the semipermeable microcapsules to render them suitable for addition to biological fluids for immunological treatment and removal of peptidic components smaller than 100,000 Daltons.
A lyophilized microcapsule obtained by a method comprising:
- (i) solubilizing antibodies in a physiological buffer comprising from about 0.3% to about 2.5% by weight alginic acid to form an antibody-containing alginic acid solution;
- (ii) introducing said antibody-containing alginic acid solution into an aqueous buffer comprising ions of a divalent salt to precipitate alginate cores, said alginate cores having diameters ranging from about 0.5 mm to about 5.0 mm and volumes ranging from about 0.1 μL to about 50 μL;
- (iii) alternately immersing said alginate cores in a polycationic macropolymer liquid buffer solutions and a polycationic macropolymer liquid buffer solution to achieve layer-by-layer electrostatic deposition and form macroion-rich liquid-liquid phases embedding said alginate cores;
- (iv) hardening said macroion-rich liquid-liquid phases by complex coacervation at ambient temperature for a period of time sufficient to form microcapsules comprising precipitated alginate cores surrounded by multilayered macroion membranes;
- (v) lyophilizing said microcapsules to produce multilayered macroion membranes that are permeable to proteins having molecular weights of up to about 100,000 Daltons to produce semipermeable microcapsules;
- (vi) conditioning said semipermeable microcapsules, and optionally classifying said microcapsules, to render them suitable for addition to biological fluids for immunological treatment and removal of peptidic components having molecular weights less than about 100,000 Daltons;
- wherein
- the polycationic macropolymers are selected from the group consisting of linear or branched polyethylenimine, polyallylamine or polyvinylamine having an average molecular weight Mn of about 100,000 to 2,000,000 Daltons, and
- the polyanionic macropolymers are selected from the group consisting of pectin (poly-D-galacturonic acid methyl ester) having a molecular weight Mn from about 20,000 to 200,000 Daltons, polyacrylic acid having a molecular weight Mn from about 10,000 to 350,000 Daltons, and carboxymethylcellulose having a molecular weight Mn from about 80,000 to 1,000,000 Daltons.
Dependencies are inferred from the claim text.