US5015576A
Expired / Lapsed
US
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Macroporous particles for cell cultivation or chromatography
Assignee: Individual
•
Filing: 1987-10-28
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Granted: 1991-05-14
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Expires: 2007-10-28
Abstract
Abstract Particles which enclose cavities can be produced by adding a water-insoluble solid, liquid or gaseous cavity generating compound to an aqueous solution of matrix material. Subsequent to forming particles by dispersion in a water-insoluble dispersion medium, the matrix is rendered insoluble in water by cooling, by covalent cross-linking or by polymerization. The cavity generating compound is washed out, whereafter the particles can be used as ion exchangers in gel filtration processes, in hydrophobic chromatography or in affinity chromatography, optionally subsequent to derivatizing the particles. The particles can also be used to advantage as microcarriers in the cultivation of anchorage-dependent cells.
Inventors
- 👤 Kjell Nilsson
- 👤 Klaus H. Mosbach
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 03:13
CPC Classifications
B; B01; B01J; B01J20/00; B01J20/28; B01J20/28002; B01J20/28004; B01J20/22; B01J20/26; B01J20/28014; B01J20/28016; B01J20/28054; B01J20/28078; B01J20/28095; B01J20/28097; C; C08; C08J; C08J3/00; C08J3/12; C08J9/00; C08J9/28; C12; C12M; C12M25/00; C12M25/16; C08J2201/00; C08J2201/04; C08J2201/054; C08J2201/0545; C08J2201/0546; Y; Y10; Y10S; Y10S435/00; Y10S435/803
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
234 words
A method for using macroporous particles in a bioreactor system for producing a biochemical product, said method comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of a water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; removing said liquid cavity generating compound from the pores of said beads to form macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; immobilizing an organism on said macroporous particles to produce an immobilized organism, said organism capable of producing said biochemical product; charging said bioreactor system with said immobilized organism; contacting said immobilized organism with a liquid reagent stream for culturing said immobilized organism; and recovering said biochemical product from said bioreactor system.
Claim 2
Independent Claim
234 words
A method for using macroporous particles in a bioreactor system for producing a biochemical product, said method comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of a water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; removing said liquid cavity generating compound from the pores of said beads to form macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; immobilizing an organism on said macroporous particles to produce an immobilized organism, said organism capable of producing said biochemical product; charging said bioreactor system with said immobilized organism; contacting said immobilized organism with a liquid reagent stream for culturing said immobilized organism; and recovering said biochemical product from said bioreactor system.
Claim 3
Dependent Claim
27 words
A method for using macroporous particles according to claim 1 wherein said bioreactor system is charged with between about 0.1 to 20 liters of said immobilized organism.
Claim 4
Dependent Claim
19 words
A method according to claim 1 wherein said organism is a hybridoma and said biochemical product comprises monoclonal antibodies.
Claim 5
Dependent Claim
21 words
A method according to claim 1 wherein said organism is an animal cell and said biochemical product comprises animal cell products.
Claim 6
Dependent Claim
23 words
A method according to claim 1 wherein said organism is a genetically engineered microbial cell and said biochemical product comprises secreted protein products.
Claim 7
Independent Claim
217 words
A method for using macroporous particles as microcarriers for culturing anchorage-dependent animal cells comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of a water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; removing said liquid cavity generating compound from the pores of said beads to form macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; mixing said macroporous particles with a growth medium and an animal cell line to form a growth support medium; incubating said growth support medium to promote cell attachment and growth; and recovering said cells from said macroporous particles.
Claim 8
Independent Claim
217 words
A method for using macroporous particles as microcarriers for culturing anchorage-dependent animal cells comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of a water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; removing said liquid cavity generating compound from the pores of said beads to form macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; mixing said macroporous particles with a growth medium and an animal cell line to form a growth support medium; incubating said growth support medium to promote cell attachment and growth; and recovering said cells from said macroporous particles.
Claim 9
Independent Claim
208 words
A method for using macroporous particles as ion-exchangers in chromatography, said method comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; and removing said liquid cavity generating compound from the pores of said beads to form said macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; derivatizing said macroporous particles to form derivatized macroporous particles having ion-exchange groups; preparing chromatography columns packed with said derivatized macroporous particles; and using said chromatography columns for ion-exchange chromatography.
Claim 10
Independent Claim
208 words
A method for using macroporous particles as ion-exchangers in chromatography, said method comprising the steps of: dissolving a water-soluble matrix material in an aqueous solvent to form an aqueous solution comprising said matrix material dissolved in said aqueous solvent; mixing a sufficient amount of water-insoluble liquid cavity generating compound with said aqueous solution of matrix material to form a dispersion of droplets of said liquid cavity generating compound in said aqueous solution; adding additional liquid cavity generating compound to said dispersion in an amount sufficient to saturate said dispersion and form droplets of said dispersion dispersed in said liquid cavity generating compound; solidifying said dispersed droplets to form beads of said matrix material having a large number of pores containing said liquid cavity generating compound dispersed therethrough; separating said solidified beads from said liquid cavity generating compound; and removing said liquid cavity generating compound from the pores of said beads to form said macroporous particles having a particle size of between about 10 to 500 micrometers and a large number of pores having diameters of between about 1-50 micrometers; derivatizing said macroporous particles to form derivatized macroporous particles having ion-exchange groups; preparing chromatography columns packed with said derivatized macroporous particles; and using said chromatography columns for ion-exchange chromatography.
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