Method and apparatus for purification of biological substances
AssigneeNone recorded
InventorHenry B. Kopf
Life of this patent
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Abstract
A process and apparatus for purifying one or more target substances from a source liquid, employing one or more cross-flow filter elements, and one or more types of chromatography resins, in combination, to provide purification with advantageous yield, product purity, and cost -and time-efficiency.
Drawings
Claims62
A process for purifying one or more target substances from a source liquid, said process comprising:
- contacting the source liquid with a chromatography resin;
- incubating the source liquid with the chromatography resin for a sufficient contact time to allow the resin to bind a desired fraction of one or more target substances,
- recirculating the chromatography resin in a cross-flow filter wherein the following steps are performed:
- concentrating the chromatography resin and separating contaminants from the chromatography-resin-bound target substance by concentration and/or diafiltration;
- eluting the target substance from the chromatography resin;
- and
- separating the target substance from the chromatography resin by diafiltration;
- recovering the target;
- and
- optionally concentrating the target substance.
A process for purifying an immunoglobulin from a source liquid, said process comprising:
- contacting the source liquid with a chromatography resin, wherein the chromatography resin comprises Protein A linked to rigid, non-porous spherical beads;
- incubating the source liquid with the chromatography resin for a sufficient contact time to allow the resin to bind a desired fraction of the immunoglobulin;
- recirculating the chromatography resin in a cross-flow filter wherein the following steps are performed:
- concentrating the chromatography resin and separating contaminants from the chromatography-resin-bound immunoglobulin by diafiltration;
- eluting the immunoglobulin from the chromatography resin;
- and
- separating the immunoglobulin from the chromatography resin by diafiltration;
- recovering the immunoglobulin;
- and
- optionally concentrating the immunoglobulin.
A method of purification of a liquid containing a target substance, comprising the steps of contacting the liquid with a chromatography resin to bind the target substance thereto, and cross-flow filtering the target substance-bound chromatography resin under elution conditions to recover a filtrate comprising the target substance.
A method of separating a liquid in a separation system comprising a bioreactor, a chromatographic resin reservoir, a first cross-flow filtration module, a second cross-flow filtration module, and a third cross-flow filtration module, said method comprising clarifying the perfusate of the bioreactor in the first cross-flow filtration module to yield a permeate, flowing the permeate to the chromatographic resin reservoir and flowing chromatographic resin and permeate to the second cross-flow filtration module for concentration, diafiltration and elution to yield an eluate, and flowing the eluate to the third cross-flow filtration module for concentration and diafiltration therein.
A method of purifying at least one target substance from a source liquid containing a bioactive contaminant, comprising the steps of:
- contacting the source liquid with a chromatography resin;
- incubating the source liquid with the chromatography resin for a sufficient contact time to bind to the chromatography resin at least one target substance from the source liquid;
- recirculating the chromatography resin in a cross-flow filter wherein the following steps are performed:
- concentration and diafiltration of the chromatography resin;
- elution of the target substance from the chromatography resin;
- and
- separation of the target substance from the chromatography resin by diafiltration;
- recovering the target substance;
- and
- optionally concentrating the target substance;
- further comprising inactivating the bioactive contaminant during said method.
A process for purifying one or more target substances from a source liquid containing said target substances together with one or more viral species, such process comprising:
- contacting the source liquid with a viral inactivation agent for the one or more viral species;
- concentrating the target substance(s) in the source liquid by tangential flow filtration in a tangential flow filter comprising a membrane effective to concentrate at least 90% by weight of the target substance(s) in such filtration;
- contacting the source liquid with a chromatography resin;
- incubating the source liquid with the chromatography resin for a sufficient contact time to bind a desired fraction of the target substance(s);
- recirculating the chromatography resin in a cross-flow filter wherein the following steps are performed:
- concentrating the chromatography resin and separating contaminants from the chromatography-resin-bound target substance by diafiltration in a filter comprising a membrane effective to retain the resin and pass species that are not bound to the resin;
- eluting the target substance from the chromatography resin by contact with an elution membrane effective to retain the resin and pass the target substance(s);
- separating the target substance from the chromatography resin by diafiltration;
- recovering the target substance(s);
- and
- optionally concentrating the target substance(s).
A process for purifying one or more target substances from a source liquid containing said target substances, such process comprising:
- contacting the source liquid with a chromatography resin;
- recirculating the chromatography resin in at least one cross-flow filter wherein the following steps are performed:
- concentrating the target substance and chromatography resin in the source liquid by tangential flow filtration;
- diafiltering the chromatography resin in a filter comprising a membrane effective to retain the resin and pass species that are not bound to the resin;
- eluting the target substance from the chromatography resin by contact with an elution membrane effective to retain the resin and pass the target substance(s);
- separating the target substance from the chromatography resin by diafiltration recovering the target substance(s);
- and
- optionally concentrating the target substance(s).
A method of manufacturing universal plasma from blood containing serological Group A and/or Group B antibodies, comprising contacting the blood containing serological Group A and/or Group B antibodies with a chromatography resin comprising corresponding Group A and/or Group B antigen, and recovering an antibodies-depleted blood product as the universal plasma.
A process for purifying a target substance from a source liquid, including the steps of contacting the source liquid with a chromatography resin having affinity for said target substance, to bind said target substance to the chromatography resin;
- and cross-flow filtering the chromatography resin to recover said target substance from the chromatography resin.
Dependencies are inferred from the claim text.