Process for producing biologically active plasminogen activator in recombinant CHO cells using suspension culture and removing detrimental components from medium
First Claim
Patent Images
1. A method of producing biologically active tissue-type plasminogen activator in a recombinant host cell suspension culture capable of producing by recombinant expression biologically active human tissue-type plasminogen activator comprising:
- a. growing recombinantly transfected Chinese hamster ovary (CHO) host cells in suspension culture using a growth supporting medium that permits the growth and maintenance of the host cells;
b. removing from the medium of said culture components in the medium that are detrimental to recovery and biological activity of said human tissue-type plasminogen activator, whilst retaining the expression capability of the host cells by a cross-flow filtration process characterized by a set of fluid-dynamic parameters utilizing a filtration apparatus having a suitable membrane such that cell culture suspension is recirculated through said filtration apparatus and a portion of the medium removed as a cell-free filtrate whilst retaining the homogeneous characteristic of said cell culture suspension;
c. replacing said filtrate with a medium devoid of components that are detrimental to recovery and activity of human tissue type plasminogen activator; and
d. collecting said biologically active human tissue-type plasminogen activator expression product present in the medium exchanged culture.
0 Assignments
0 Petitions
Accused Products
Abstract
A method of producing biologically active human tissue plasminogen activator in suspension culture is provided wherein recombinant Chinese hamster ovary cells are cultured and certain components detrimental to recovery and biological activity are removed as a cell-free filtrate by cross-flow filtration. The human tissue plasminogen activator is recovered from the culture medium.
-
Citations
8 Claims
-
1. A method of producing biologically active tissue-type plasminogen activator in a recombinant host cell suspension culture capable of producing by recombinant expression biologically active human tissue-type plasminogen activator comprising:
-
a. growing recombinantly transfected Chinese hamster ovary (CHO) host cells in suspension culture using a growth supporting medium that permits the growth and maintenance of the host cells; b. removing from the medium of said culture components in the medium that are detrimental to recovery and biological activity of said human tissue-type plasminogen activator, whilst retaining the expression capability of the host cells by a cross-flow filtration process characterized by a set of fluid-dynamic parameters utilizing a filtration apparatus having a suitable membrane such that cell culture suspension is recirculated through said filtration apparatus and a portion of the medium removed as a cell-free filtrate whilst retaining the homogeneous characteristic of said cell culture suspension; c. replacing said filtrate with a medium devoid of components that are detrimental to recovery and activity of human tissue type plasminogen activator; and d. collecting said biologically active human tissue-type plasminogen activator expression product present in the medium exchanged culture. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
-
Specification