Purification of biological substances
First Claim
Patent Images
1. 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 diafiltration;
eluting the target substance from the chromatography resin; and
separating the target substance from the chromatography resin by diafiltration;
recovering the target substance; and
optionally concentrating the target substance, wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands, bound thereto.
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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.
70 Citations
70 Claims
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1. A process for purifying one or more target substances from a source liquid, said process comprising:
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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 diafiltration;
eluting the target substance from the chromatography resin; and
separating the target substance from the chromatography resin by diafiltration;
recovering the target substance; and
optionally concentrating the target substance, wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands, bound thereto. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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- 22. 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, wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands bound thereto.
- 43. 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, wherein the chromatographic resin comprises porous chromatographic resin particles that are functionalized with affinity ligands bounds thereto.
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47. A method of purifying at least one target substance from a source liquid containing a bioactive contaminant, comprising the steps of:
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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 wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands, bounds thereto. - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60)
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61. 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:
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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); and
separating the target substance from the chromatography resin by diafiltration;
recovering the target substance(s); and
optionally concentrating the target substance(s), wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands, bounds thereto. - View Dependent Claims (62, 63)
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64. A process for purifying one or more target substances from a source liquid containing said target substances, such process comprising:
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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); and
separating the target substance from the chromatography resin by diafiltration;
recovering the target substance(s); and
optionally concentrating the target substance(s), wherein the chromatography resin comprises porous chromatography resin particles that are functionalized with affinity ligands bounds thereto. - View Dependent Claims (65, 66, 67, 68, 69, 70)
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Specification