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G. Baklayan, B.A. Aird, T.W. Joe, J. Yang, T.L. Thetford, E.C. Collins; Comparison of the Purity of Various Bovine and Ovine Testicular Hyaluronidase Preparations to a Preparation of Purified Ovine Testicular Hyaluronidase (Vitrase®) . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5105.
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© ARVO (1962-2015); The Authors (2016-present)
To compare various commercial bovine (BH) and ovine (OH) testicular hyaluronidase preparations for purity.
Vitrase® (hyaluronidase ovine for injection) is the only FDA approved, highly purified, preservative–free, ovine hyaluronidase preparation (ISTA Pharmaceuticals, Inc.). A marketed bovine product, Amphadase® (Amphastar Pharmaceuticals, Inc.), was also obtained for inclusion in a separate, but identical analysis. Vitrase, Amphadase and other commercially available BH and OH testicular hyaluronidase preparations (Sigma, CP Pharmaceuticals), were analyzed and compared for purity utilizing sodium dodecyl (lauryl) sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and Western Blots. The bovine hyaluronidase from Biozyme, HY–B6, underwent an identical purification process to Vitrase.
Equivalent amounts of protein samples of BH and OH preparations were analyzed by SDS–PAGE. In the molecular weight range of 50 – 90 kDa, there were prominent bands in all hyaluronidase preparations. This is the range where hyaluronidase and any large molecular weight fragments (breakdown products) of hyaluronidase will appear. Bands in the migration positions of Annexin II and IgG fragment are also generally present in all hyaluronidase preparations, although in less pure materials, the bands are less distinct.
Overall, the BH preparations appear to be less pure than the OH preparations including the Amphadase BH product. The Biozyme BH preparation, HY–B6, seems to be the one exception and its gel pattern appears to be similar to Vitrase.
There were differences in the purity of available OH and BH preparations. Vitrase appears to be the purest drug product commercially available. The Biozyme BH preparation, HY–B6, underwent the same purification process as Vitrase and showed similar purity.
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