Claims for Patent: 6,136,960

Summary for Patent: 6,136,960

Title:	Method for evaluation of the ratio of amounts of biomolecules or their sub-populations in a mixture
Abstract:	The present invention is directed to a method of determining the ratio of the concentration or amounts of two or more chemical compounds and/or biopolymers, or of the relative amounts of sub-populations of closely structurally related compounds, in a multi-component mixture, extract, system, and the like using an analytical physicochemical process. The method is based on distribution of the components of the mixture (system, extract, etc.) between two or more immiscible phases and the subsequent determination of the total amounts (concentrations) of biopolymers (chemical compounds) or of the amounts (concentrations) of one or more component(s) or sub-populations of the system in the phases. The ratio between these concentrations is defined therein as the partition coefficient. The partition coefficient is used as a measure of the ratio of the amounts of two or more biopolymers (chemical compounds) or their sub-populations in a multi-component mixture (extract, system, etc.), when calibrated against a series of sample systems of the same type with varied ratios of components.
Inventor(s):	Chait; Arnon (Bay Village, OH), Zaslavsky; Boris Y. (Solon, OH)
Assignee:	Zaslavsky; Boris Y. (Solon, OH) Chait; Arnon (Bay Village, OH)
Application Number:	09/135,751
Patent Claims:	1. A method for the quantitative analysis of a multi- component mixture wherein the method of quantitative analysis comprises the steps of: i) forming a partitioning system capable of separation into two or more immiscible phases, ii) mixing a sample of the mixture to be quantitatively analyzed with the partitioning system; iii) maintaining the partitioning system of step (ii) until phase separation occurs; iv) measuring the total amount or total concentration of components in the mixture, or of a single class of components within said mixture, in each of the separated phases wherein each of the components being analyzed in the mixture exists in each of the separated phases; v) determining the overall partition coefficient (K) of the mixture, or of the single class of components within the mixture, wherein the overall partition coefficient (K) of the mixture is defined as the ratio between the amount or concentrations of the mixture, or the single class of components within the mixture, in each of the separated phases; and, vi) comparing the determined overall partition coefficient (K) of the mixture, or the single components within the mixture, with a range of overall partition coefficients obtained from known reference mixtures of the same components thereby assessing the ratio of the particular components of the mixture under characterization without the need to pre-separate the components. 2. The method of claim 1 wherein the pH in the mixture under analysis is within the range of pH from 1.0 to 11.0. 3. The method of claim 1 wherein the phases are of aqueous nature. 4. The method of claim 1 wherein the total weight of the initial mixture being partitioned does not exceed 20 % wt. of the total two-phase system weight. 5. The method of claim 1 wherein the partition coefficient of the overall mixture is measured with a total assay technique wherein said assay techniques comprise dye-based assays, organic nitrogen technique, direct spectrophotometry, and derivatization-based techniques. 6. The method of claim 1 wherein the partition coefficient of a single class of components within the mixture is measured with immunoassay, enzyme activity assay, or biospecific assay. 7. The method of claim 1 wherein the overall partition coefficient of a single class of components within the mixture is used as a measure of the ratio of the amount or concentration of the single class of components of the mixture. 8. The method of claim 1 wherein the temperature of the partitioning system is in the range of 4.degree. C. to 60.degree. C. 9. The method of claim 1 wherein the composition of the partitioning system provides a quantitative relationship between the overall partition coefficient of the mixture and the ratio between the amount or concentration of the components of interest. 10. The method of claim 1 wherein the partition coefficient is measured for the sample of a said mixture in two or more aqueous two-phase systems of different compositions selected from the group consisting of a polymer/polymer system and a polymer/salt system. 11. The method of claim 1 wherein the partitioning system used for partitioning of the mixture is calibrated with partition coefficients (K) for a specifically selected set of low and high molecular weight compounds in addition to the reference samples. 12. The method of claim 1 wherein the overall partition coefficient of the multi-component mixture is used as an indicator of changes in the ratios of the amount or concentration of the components or as an indicator of differences in the multi-component mixture being analyzed relative to those in the mixture used as a reference standard. 13. The method of claim 1 wherein the multi-component mixture is a protein mixture. 14. The method of claim 13 wherein the protein mixture is selected from the group consisting of lysosome/concanavalin A, serum albumin/human .gamma.-globulin, lysosome/hemoglobin, myoglobin/.beta.-lactoglobulin, hemoglobin/glycated hemoglobin, transferrin/carbohydrate-deficient transferrin and Z'-deoxyadenosine/adenosine 5'-monophosphate sodium salt protein mixtures. 15. The method of claim 1 wherein the multi-component mixture is a mixture of biopolymers or sub-populations of said biopolymers.

Details for Patent 6,136,960

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2016-04-19
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2016-04-19
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2016-04-19