APPARATUS AND PROCESS FOR PURIFICATION OF PROTEINS
The invention is directed to an apparatus and method for purifying a protein. The apparatus involves the use of a capture chromatography resin, a depth filter arranged after the capture chromatography resin, and a mixed-mode chromatography resin arranged after the depth filter. The method involves providing a sample containing the protein, processing the sample through a capture chromatography resin, a depth filter, and a mixed-mode chromatography resin. A membrane adsorber or monolith may be substituted for the mixed-mode chromatography column.
1 - 20 . (canceled)
21 . A method for purifying a protein comprising:
a. providing a sample containing the protein;
b. processing the sample through a capture chromatography resin to provide a first eluate comprising the protein;
c. after the sample is processed through the capture chromatography resin, processing the first eluate through a depth filter to provide a filtered eluate comprising the protein; and
d. after the first eluate is processed through the depth filter, processing the filtered eluate through a mixed-mode chromatography resin to provide a second eluate comprising the protein.
22 . The method of claim 21 wherein the capture chromatography resin is selected from the group consisting of an affinity resin, an ion exchange resin, and a hydrophobic interaction resin.
23 . The method of claim 21 wherein the capture chromatography resin is selected from the group consisting of a protein A resin, a protein G resin, a protein A/G resin, and a protein L resin.
24 . The method of claim 21 wherein the protein is selected from the group consisting of a protein fragment, an antibody, a monoclonal antibody, an immunoglobulin, and a fusion protein.
25 . The method of claim 21 wherein the sample is a cell culture.
26 . The method of claim 21 wherein the sample is clarified prior to processing through the capture chromatography resin.
27 . The method of claim 26 wherein the sample is clarified by a clarification method selected from the group consisting of centrifugation, microfiltration, ultrafiltration, depth filtration, sterile filtration, and treatment with a detergent.
28 . The method of claim 21 wherein the first eluate is subjected to viral inactivation after processing through the capture chromatography resin but before processing through the depth filter.
29 . The method of claim 28 wherein the viral inactivation comprises a method selected from the group consisting of treatment with acid, detergent, chemicals, nucleic acid cross-linking agents, ultraviolet light, gamma radiation, and heat.
30 . The method of claim 21 wherein the filtered eluate is processed through a depth filter a second time.
31 . The method of claim 21 wherein the mixed-mode chromatography resin comprises a chromatography resin utilizing one or more chromatography techniques selected from the group consisting of anion exchange, cation exchange, hydrophobic interaction, hydrophilic interaction, hydrogen bonding, pi-pi bonding, and metal affinity.
32 . The method of claim 21 wherein the mixed-mode chromatography resin comprises a chromatography resin utilizing a combination of anion exchange and hydrophobic interaction chromatography techniques.
33 . The method of claim 21 wherein, after processing through the mixed-mode chromatography resin, the second eluate is subjected to further filtration.
34 . The method of claim 33 wherein the further filtration comprises one or more of the methods selected from the group consisting of viral filtration, nanofiltration, ultrafiltration, and diafiltration.
35 . The method of claim 21 wherein filtered eluate is processed through the mixed-mode chromatography resin in flow-through mode.
36 . The method of claim 21 wherein filtered eluate is processed through the mixed-mode chromatography resin in bind-elute mode.
37 . A method for purifying a protein comprising:
a. providing a sample containing the protein;
b. processing the sample through a capture chromatography resin to provide a first eluate comprising the protein;
c. after the sample is processed through the capture chromatography resin, processing the first eluate through a depth filter to provide a filtered eluate comprising the protein; and
d. after the first eluate is processed through the depth filter, processing the filtered eluate through a membrane adsorber to provide a second eluate comprising the protein.
38 . A method for purifying a protein comprising:
a. providing a sample containing the protein;
b. processing the sample through a capture chromatography resin to provide a first eluate comprising the protein;
c. after the sample is processed through the capture chromatography resin, processing the first eluate through a depth fitter to provide a filtered eluate comprising the protein; and
d. after the first eluate is processed through the depth filter, processing the filtered eluate through a monolith to provide a second eluate comprising the protein.