Analysis and separation of platelet-derived growth factor...

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

Reexamination Certificate

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C530S416000, C530S399000, C435S069100, C435S254210

Reexamination Certificate

active

07084262

ABSTRACT:
Methods for improving purification and quantification of platelet derived growth factor (PDGF) proteins having structural heterogeneity are provided. Preparation of substantially pure isoforms of these proteins is achieved using TSK sulfopropyl cation exchange chromatography and reverse phase high performance liquid chromatography. A reverse charged capillary zone electrophoresis method enables quantification of substantially pure isoforms of these proteins resulting from endoproteolytic post-translational modifications. Compositions of the invention are substantially purified isoforms of secreted PDGF proteins having structural heterogeneity, more particularly purified intact, single-clipped, and double-clipped isoforms of recombinant PDGF-BB. Pharmaceutical compositions comprising at least one of these substantially purified recombinant PDGF isoforms and methods for their use in promoting wound healing are also provided.

REFERENCES:
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Soma et al., FASEB Journal: official publication of the Federation of American Societies for Experimental Biology, (Aug. 1992) 6(11) 2996-3001.
Cook, A.L., et al., Purification and Analysis of Proteinase-Resistant Mutants of Recombinant Platelet-Derived Growth Factor-BB Exhibiting Improved Biological Activity,Biochemical Journal, 1992, pp. 56-65, vol. 281(1).
Oefner, C., et al., “Crystal Structure of Human Platelet-Derived Growth Factor BB,”EMBO Journal, 1992, pp. 3921-3926, vol. 11.

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