Preparation of human placental hyaluronidase
AssigneeThe Green Cross Corporation
InventorsYasuhiro Mima, Masaaki Yamada
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Abstract
Human placental hyaluronidase preparation is prepared by extracting hyaluronidase from human placenta with an aqueous alkaline solution, harvesting globulin fraction from the extract, selectively adsorbing hyaluronidase contained in the fraction on an anion exchanger such as DEAE-Cellulose and eluting it, gel-filtrating the eluate, and lyophilizing the filtrate into powder. The hyaluronidase thus obtained has a molecular weight of about 70,000, with isoelectric point of pH 5.2, and enzymatic optical pH ranging 3.6 - 4.0, differing from conventional bovin testicular hyaluronidase. The preparation contains no heterogeneous proteins which cause ill-effect to human body.
Claims14
A human placental hyaluronidase preparation free from hypotensive factors, blood type substances, and hemoprotein, said human placental hyaluronidase having a molecular weight of about 70,000, an isoelectric point of pH about 5.2 and enzymatic optimum pH ranging from 3.6 to 4.0, and being stable for at least 24 hours when kept below 30.degree. C. at pH 6-7 in an aqueous solution.
A process for preparing a hyaluronidase preparation comprising subjecting a human placental tissue to extraction with an aqueous alkaline solution at a pH of about 7.5 to about 10.5, harvesting globulin fraction from the extract, contacting the globulin fraction with an anion exchanger equilibrated with about 0.005 M to about 0.01 M buffer at a pH of about 6.0 to about 8.5 and eluting hyaluronidase adsorbed with a buffer having an increased molarity and/or a lowered pH, and subjecting thus obtained hyaluronidase fraction to gel-filtration.
Dependencies are inferred from the claim text.