Virus like particle purification
AssigneeTakeda Vaccines (Montana), Inc.
InventorsThomas S. Vedvick, Bryan Steadman, Charles Richardson, Thomas R. Foubert, Charles R. Petrie
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Abstract
Methods for purifying human Calciviruses are disclosed, including Noroviruses and Sapoviruses.
Claims29
A method of purifying Calicivirus virus-like particles (VLPs) using a multistep chromatographic process, wherein said VLPs are produced in insect cells, said method comprising
- (a) contacting a cell lysate or culture supernatant containing said VLPs with an ion exchange chromatographic material, wherein said VLPs bind to said ion exchange chromatographic material;
- (b) contacting the VLPs eluted from said ion exchange chromatographic material with a hydrophobic interaction chromatographic material, wherein said VLPs bind to said hydrophobic interaction chromatographic material;
- and
- (c) eluting said VLPs from said hydrophobic interaction chromatographic material, wherein said VLPs are purified to greater than about 70%,
- wherein the chromatographic materials used in the method do not comprise hydroxyapatite.
A method of purifying Calicivirus virus-like particles (VLPs) wherein the VLPs are produced in insect cells, said method comprising contacting a solution containing said VLPs with two or more chromatographic materials, wherein the chromatographic materials do not comprise hydroxyapatite, and wherein at least one chromatographic material retains said VLPs.
A method of purifying Calicivirus virus-like particles (VLPs) using a multistep chromatographic process wherein said VLPs are produced in insect cells, said method comprising
- (a) contacting a cell lysate or culture supernatant containing said VLPs with an ion exchange chromatographic material, wherein said VLPs bind to said ion exchange chromatographic material, and wherein said ion exchange chromatographic material comprises an anion exchanger;
- (b) contacting the VLPs eluted from said ion exchange chromatographic material with a hydrophobic interaction chromatographic material, wherein said VLPs bind to said hydrophobic interaction chromatographic material;
- and
- (c) eluting said VLPs from said hydrophobic interaction chromatographic material, wherein said VLPs are purified to greater than about 70%,
- wherein the chromatographic materials used in the method do not comprise hydroxyapatite.
A method of purifying Calicivirus virus-like particles (VLPs) using a multistep chromatographic process wherein said VLPs are produced in insect cells, said method comprising
- (a) contacting a cell lysate or culture supernatant containing said VLPs with an ion exchange chromatographic material, wherein said VLPs bind to said ion exchange chromatographic material, and wherein said cell lysate or culture supernatant is filtered, centrifuged, or clarified prior to contact with the ion exchange chromatographic material;
- (b) contacting the VLPs eluted from said ion exchange chromatographic material with a hydrophobic interaction chromatographic material, wherein said VLPs bind to said hydrophobic interaction chromatographic material;
- and
- (c) eluting said VLPs from said hydrophobic interaction chromatographic material, wherein said VLPs are purified to greater than about 70%,
- wherein the chromatographic materials used in the method do not comprise hydroxyapatite.
A method of purifying Calicivirus virus-like particles (VLPs) using a multistep chromatographic process wherein said VLPs are produced in insect cells, said method comprising
- (a) contacting a cell lysate or culture supernatant containing said VLPs with an ion exchange chromatographic material, wherein said VLPs bind to said ion exchange chromatographic material, and wherein said cell lysate or culture supernatant is clarified by precipitation or by size exclusion chromatography;
- (b) contacting the VLPs eluted from said ion exchange chromatographic material with a hydrophobic interaction chromatographic material, wherein said VLPs bind to said hydrophobic interaction chromatographic material;
- and
- (c) eluting said VLPs from said hydrophobic interaction chromatographic material, wherein said VLPs are purified to greater than about 70%,
- wherein the chromatographic materials used in the method do not comprise hydroxyapatite.
Dependencies are inferred from the claim text.