Abstract: Mutant Proteus vulgaris strains are provided that, when grown in the absence of an exogenous chondroitinase I and II inducer, produce P. vulgans chondroitinase I and chondroitinase II proteins. The mutants typically produce chondroitinase I and II proteins in the absence of exogenous inducers and in amounts in excess of those produced by wild-type P. vulgaris strains induced with such inducers. Two classes of such mutants, Classes 1 and 2, are disclosed. Class 1 and class 2 mutants differ in the relative amounts of chondroitinases I and II produced when cells are grown in casamino acids--supplemented minimal medium. Additional phenotypic variants that release chondroitinase I protein into the culture medium are provided as well. Also contemplated is a method for producing P. vulgaris chondroitinase I and II proteins.

Abstract: Chondroitinase II, a protein having an isoelectric point of approximately 8.4-8.45 and an apparent molecular mass of 112 kDa when electrophoresed in a 4 to 20% gradient acrylamide gel in 25 mM Tris/192 mM glycine buffer at pH 8.5 in the presence of about 0.1% (w/v) SDS, has been isolated and purified. A process for the copurification by affinity chromatography of the chondroitinase I and chondroitinase II proteins produced by Proteus vulgaris is also provided. The proteins can be further purified by metal chelating chromatography. Therapeutic or surgical compositions of isolated chondroitinases I and II or the copurified mixture of chondroitinase I and II are also disclosed. These compositions are used in a method for selectively and completely disinserting the vitreous body from the neural retina of an eye.

Abstract: Chondroitinase II, a protein having an isoelectric point of approximately 8.4-8.45 and an apparent molecular mass of 112 kDa when electrophoresed in a 4 to 20% gradient acrylamide gel in 25 mM Tris/192 mM glycine buffer at pH 8.5 in the presence of about 0.1% (w/v) SDS, has been isolated and purified. A process for the copurification by affinity chromatography of the chondroitinase I and chondroitinase II proteins produced by Proteus vulgaris is also provided. The proteins can be further purified by metal chelating chromatography. Therapeutic or surgical compositions of isolated chondroitinases I and II or the copurified mixture of chondroitinase I and II are also disclosed. These compositions are used in a method for selectively and completely disinserting the vitreous body from the neural retina of an eye.

Abstract: Chondroitinase II, a protein having an isoelectric point of approximately 8.4-8.45 and an apparent molecular mass of 112 kDa when electrophoresed in a 4 to 20% gradient acrylamide gel in 25 mM Tris/192 mM glycine buffer at pH 8.5 in the presence of about 0.1% (w/v) SDS, has been isolated and purified. A process for the copurification by affinity chromatography of the chondroitinase I and chondroitinase II proteins produced by Proteus vulgaris is also provided. The proteins can be further purified by metal chelating chromatography. Therapeutic or surgical compositions of isolated chondroitinases I and II or the copurified mixture of chondroitinase I and II are also disclosed. These compositions are used in a method for selectively and completely disinserting the vitreous body from the neural retina of an eye.

Abstract: This invention relates to methods for the isolation and purification of the recombinantly expressed major protein component of chondroitinase ABC, which is referred to as "chondroitinase I", from Proteus vulgaris (P. vulgaris). This invention further relates to methods for the isolation and purification of the recombinantly expressed second protein component of chondroitinase ABC, which is referred to as "chondroitinase II", from P. vulgaris. These methods provide significantly higher yields and purity than those obtained by adapting for the recombinant enzymes the method previously used for isolating and purifying native chondroitinase I enzyme from P. vulgaris.

Abstract: A chromatographic process for the copurification of chondroitinase proteins useful in ocular surgery for non-surgical disruption of chondroitin sulfate, the molecule which mediates the attachment between the retina and vitreous body in the human eye. The process involves the use of ion exchange resins in conjunction with an affinity elution with chondroitin sulfate to afford copurification of the proteins.

Abstract: A chondroitinase II is isolated from Proteus vulgaris. This enzyme together with chondroitinase I, is useful for selectively and completely disinserting the ocular vitreous body from the neural retina of the eye. The chondroitinase II has the amino acid sequence of SEQ. ID No. 2, an isoelectric point of from about 8.4 to about 8.45, and a molecular weight of 111,772+27 daltons as determined by electrospray and 111,725+20 daltons as determined by laser desorption. Preferrably, the chondroitinase II and chondroitinase I are co-purified chondroitinases which can then be separately eluted from each other.