Copolymer 1 related polypeptides for use as molecular weight markers and for therapeutic use
AssigneeYeda Research and Development Co. Ltd.
InventorsAlexander Gad, Dora Lis
Life of this patent
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Abstract
The present invention provides processes for determining the molecular weight of glatiramer acetate and other copolymers using molecular weight markers. The present invention further provides a plurality of molecular weight markers for determining the molecular weight of glatiramer acetate and other copolymers which display linear relationships between molar ellipticity and molecular weight, and between retention time and the log of the molecular weight. The molecular weight markers also optimally demonstrate biological activity similar to glatiramer acetate or corresponding copolymers and can be used for treating or preventing various immune diseases.
Claims26
In a process for obtaining a pharmaceutical product containing a mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine, wherein the mixture has an average molecular weight from 4000 to 13,000 Daltons and in the mixture the molar fraction of alanine is 0.427, of glutamic acid is 0.141, of lysine is 0.337 and of tyrosine is 0.093 and wherein the process includes determining the molecular weight distribution of a batch of an aqueous mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine, using a gel permeation chromatography column to determine whether the mixture has an average molecular weight from 4000 to 13,000 Daltons for inclusion in the pharmaceutical product, the improvement comprising
- calibrating the molecular weight obtained using the gel permeation chromatography column by subjecting a plurality of molecular weight markers, each of which is a polypeptide consisting essentially of alanine, glutamic acid, tyrosine and lysine and having a predetermined amino acid sequence, to chromatography on the column to establish a relationship between retention time on the column and molecular weight.
A process for obtaining a pharmaceutical product containing a mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine, wherein the mixture has an average molecular weight from 4000 to 13,000 Daltons and in the mixture the molar fraction of alanine is 0.427, of glutainic acid is 0.141, of lysine is 0.337 and of tyrosine is 0.093, which comprises obtaining a batch of a mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine;
- determining the average molecular weight of the mixture of polypeptides in the batch using a molecular weight-calibrated gel permeation chromatography column;
- and
- including in the pharmaceutical product the mixture if the mixture is determined to have an average molecular weight from 4000 to 13,000 Daltons,
- wherein the gel permeation chromatography column is calibrated by subjecting a plurality of molecular weight markers to chromatography on the column to establish a relationship between the retention time on the column and molecular weight, wherein each of the markers is a polypeptide consisting essentially of alanine, glutamic acid, tyrosine and lysine and has a predetermined amino sequence.
A process for determining the average molecular weight of an aqueous mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine, wherein in the mixture the molar fraction of alanine is 0.427, of glutamic acid is 0.141, of lysine is 0.337 and of tyrosine is 0.093, which comprises subjecting the mixture to chromatography on a molecular weight-calibrated gel permeation chromatography column so as to determine the average molecular weight of the mixture, wherein the gel permeation chromatography column is calibrated by subjecting a plurality of molecular weight markers to chromatography on the column to establish a relationship between retention time on the column and molecular weight, wherein each of the markers is a polypeptide consisting essentially of alanine, glutamic acid, tyrosine and lysine and has a predetermined amino acid sequence.
A process for determining whether an aqueous mixture of polypeptides, each of which consists essentially of alanine, glutamic acid, tyrosine and lysine, has an average molecular weight from 4000 to 13,000 Daltons, wherein in the mixture the molar fraction of alanine is 0.427, of glutamic acid is 0.141, of lysine is 0.337 and of tyrosine is 0.093, which process comprises subjecting the mixture to a calibrated gel permeation chromatography column to determine the average molecular weight of the mixture wherein the gel permeation chromatography column is calibrated by subjecting a plurality of molecular weight markers to chromatography on the column to establish a relationship between retention time on the column and molecular weight, wherein each of the markers is a polypeptide consisting essentially of alanine, glutamic acid, tyrosine and lysine and has a predetermined amino acid sequence.
Dependencies are inferred from the claim text.