Back to results
US 2023/0416342 A1Published application
Methods for purifying inter-alpha inhibitor proteins
AssigneeProThera Biologics, Inc.
InventorsYow-Pin LIM, Joseph QIU
Life of this patent
Estimating the term…
Abstract
Described herein are methods for purifying IαIp from a biological material using an endotoxin-binding agent. The method involves applying the biological material containing the IαIp to an endotoxin-binding agent, discarding the flow through, applying a wash buffer(s), and eluting the IαIp from the endotoxin-binding agent.
Claims149
A method of purifying an inter-alpha inhibitor protein (IαIp) from a biological material comprising:
- (a) applying the biological material comprising the IαIp to an endotoxin-binding agent and separating a flow through comprising the biological material that does not bind to the endotoxin-binding agent;
- and
- (b) applying an elution buffer comprising a salt to the endotoxin-binding agent and collecting an eluate comprising the IαIp.
A method of purifying an IαIp from plasma comprising:
- (a) diluting the plasma with a dilution buffer comprising deionized water to form diluted plasma;
- (b) applying the diluted plasma to an ETOXICLEAR™ resin and separating a flow through comprising the diluted plasma that does not bind to the ETOXICLEAR™ resin;
- (c) applying a first wash buffer comprising about 75 mM glycine, about 100 mM AcOH, and about 150 mM NaCl at a pH of about 5.2 to the ETOXICLEAR™ resin and separating a flow through comprising the first wash buffer;
- (d) applying a second wash buffer comprising about 20 mM Tris-HCl and about 300 mM NaCl at a pH of about 7.2 to the ETOXICLEAR™ resin and separating a flow through comprising the second wash buffer;
- and
- (e) applying an elution buffer comprising about 20 mM Tris-HCl and about 500 mM NaCl at a pH of 7.2 to the ETOXICLEAR™ resin and collecting an eluate comprising the IαIp.
3 more elements
A method of purifying an IαIp from plasma comprising:
- (a) diluting the plasma with a dilution buffer comprising 15 mM phosphate at a pH of about 5.5 to the plasma to form diluted plasma;
- (b) applying the diluted plasma to a DETOXI-GEL™ resin and separating a flow through comprising the diluted plasma that does not bind to the DETOXI-GEL™ resin;
- (c) applying a first wash buffer comprising about 15 mM phosphate and about 50 mM NaCl at a pH of about 5.5 to the DETOXI-GEL™ resin and separating a flow through comprising the first wash buffer;
- (d) applying a second wash buffer comprising about 15 mM phosphate and about 100 mM NaCl at a pH of about 5.5 to the DETOXI-GEL™ resin and separating a flow through comprising the second wash buffer;
- and
- (e) applying an elution buffer comprising about 15 mM phosphate and about 1,000 mM NaCl at a pH of about 5.5 to the DETOXI-GEL™ resin and collecting an eluate comprising the IαIp.
3 more elements
Dependencies are inferred from the claim text.