Process for production and purification of recombinant lysosomal alpha-mannosidase
AssigneeZymenex A/S
InventorsJens Fogh, Claes Andersson, Cecilia Weigelt, Pia Hydén, Helena Reuterwall, Stefan Nilsson
Life of this patent
Estimating the term…
Abstract
The present invention relates to a process for purification of recombinant alpha-mannosidase, a process for production of alpha-mannosidase, a composition comprising alpha-mannosidase, use of the composition as a medicament, use as a medicament for the treatment of alpha-mannosidosis and a method of treating alpha-mannosidosis and/or alleviating the symptoms of alpha-mannosidosis.
Claims11
(canceled)
A composition comprising alpha-mannosidase obtainable by a process for purification of a recombinant human lysosomal alpha-mannosidase from a cell culture, wherein a fraction of said cell culture comprising said recombinant human lysosomal alpha-mannosidase is subjected to chromatography on a resin comprising a multi-modal ligand, wherein said resin bound multi-modal ligand is a substance having a carboxylic acid or sulphonic acid group.
A process for fed batch or continuous production of recombinant alpha-mannosidase, comprising the following steps:
- a. inoculating a production reactor comprising a base medium with cells capable of producing recombinant alpha-mannosidase on day 0, to provide a cell culture;
- b. adding a feed medium to said cell culture at least once from day 1;
- c. adjusting the temperature of said cell culture to at the most 35° C., either after day 3 or when the viable cell density is higher than 2.1 MVC/mL, whichever comes first;
- and
- d. applying a process for purification of the recombinant human lysosomal alpha-mannosidase from the cell culture, wherein a fraction of said cell culture comprising said recombinant human lysosomal alpha-mannosidase is subjected to chromatography on a resin comprising a multi-modal ligand, wherein said resin bound multi-modal ligand is a substance having a carboxylic acid or sulphonic acid group.
A composition comprising purified recombinant alpha-mannosidase, wherein at least 80% of the alpha-mannosidase is present as a 130 kDa glycoprotein.
A method of treating alpha-mannosidosis and/or reducing or alleviating the symptoms associated with alpha-mannosidosis, said method comprising a step of administering a composition comprising:
- A) a composition comprising alpha-mannosidase obtainable by a process for purification of a recombinant human lysosomal alpha-mannosidase from a cell culture, wherein a fraction of said cell culture comprising said recombinant human lysosomal alpha-mannosidase is subjected to chromatography on a resin comprising a multi-modal ligand, wherein said resin bound multi-modal ligand is a substance having a carboxylic acid or sulphonic acid group;
- B) a composition comprising alpha-mannosidase obtainable by a process for fed batch or continuous production of recombinant alpha-mannosidase, comprising the following steps:
- i. inoculating a production reactor comprising a base medium with cells capable of producing recombinant alpha-mannosidase on day 0, to provide a cell culture;
- ii. adding a feed medium to said cell culture at least once from day 1;
- iii. adjusting the temperature of said cell culture to at the most 35° C., either after day 3 or when the viable cell density is higher than 2.1 MVC/mL, whichever comes first;
- and
- iv. applying a process for purification of the recombinant human lysosomal alpha-mannosidase from the cell culture, wherein a fraction of said cell culture comprising said recombinant human lysosomal alpha-mannosidase is subjected to chromatography on a resin comprising a multi-modal ligand, wherein said resin bound multi-modal ligand is a substance having a carboxylic acid or sulphonic acid group;
- or
- C) a composition comprising purified recombinant alpha-mannosidase, wherein at least 80% of the alpha-mannosidase is present as a 130 kDa glycoprotein.
Dependencies are inferred from the claim text.