Compositions and methods for purifying calreticulin
AssigneeCalretex, LLC
InventorsMarek Michalak, Monika Dabroska
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Abstract
The present invention relates to compositions, methods for expressing, and related methods for purifying calreticulin that is free of an affinity label or tag (i.e., non-tagged calreticulin). The invention provides useful methods for commercial production of human calreticulin in a bacterial expression system such as Escherichia coli.
Claims30
A method for producing calreticulin comprising:
- (a) culturing a microorganism having a calreticulin gene under suitable culture conditions;
- (b) inducing expression of the calreticulin gene to produce a calreticulin protein product that is free of an affinity tag or label;
- and (c) purifying the calreticulin product.
A method for expressing calreticulin in a prokaryotic cell, which comprises:
- (a) culturing a prokaryotic cell comprising a vector under conditions conducive to gene expression, wherein the vector comprises a nucleic acid encoding calreticulin, an origin of replication, and at least one selectable marker, and wherein the nucleic acid is operatively associated with an expression control sequence;
- (b) inducing expression of the calreticulin gene to produce a calreticulin protein product that is free of an affinity tag or label;
- and
- (c) purifying said calreticulin protein product.
A vector comprising an origin of replication, at least one selectable marker, and a multiple cloning site comprising a nucleic acid encoding a signal sequence and calreticulin, and wherein said nucleic acid is operatively associated with an expression control sequence, and wherein said nucleic acid encodes a termination signal.
Dependencies are inferred from the claim text.