Concanavalin a, methods of expressing, purifying and characterizing concanavalina, and sensors including the same
First Claim
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1. A composition comprising a substantially purified lectin polypeptide wherein the composition is at least 95% pure.
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Abstract
A novel method for purifying various lectins is disclosed. More specifically a novel method for purifying Concanavalin A is set forth. Methods of expressing purifying and characterizing a mutant Concanavalin A, and sensors including the foregoing are also disclosed.
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43 Claims
- 1. A composition comprising a substantially purified lectin polypeptide wherein the composition is at least 95% pure.
- 2. A composition comprising a substantially purified lectin polypeptide wherein the lectin comprises greater than 95% by weight of the total protein of the composition.
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3. A composition comprising a substantially purified lectin polypeptide wherein the composition has a purity of greater than 95% as determined by relative peak area integration.
- 11. A method of producing a recombinant lectin of interest comprising inducing expression of said lectin in a bacterial cell culture.
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22. A method of purifying a lectin comprising:
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adding a denaturing, chaotropic agent to a solution of lectin having a pH less than 5, and subjecting said solution to size exclusion chromatography. - View Dependent Claims (23)
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- 24. A composition comprising a substantially purified lectin having less than about 150 ng of Host Cell Protein (HCP) per mg of purified lectin.
- 28. An isolated nucleic acid sequence encoding a mutant form of a natural Concanavalin A.
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34. A method of producing a Concanavalin A exhibiting reduced precipitation during purification comprising performing a mutation to the nucleic acid sequence of a Concanavalin A wherein the mutation encodes for an amino acid change, the amino acid change converting an acidic amino acid site to a neutral amino acid.
- 35. A vector comprising an inducible promoter, a kanamycin resistance gene and a nucleic acid sequence encoding for a form of Concanavalin A.
- 37. A sensor comprising a mutant form of Concanavalin A.
Specification