US 9,150,626 B2 · Protein surface remodeling

Abstract

Aggregation is a major cause of the misbehavior of proteins. A system for modifying a protein to create a more stable variant is provided. The method involves identifying non-conserved hydrophobic amino acid residues on the surface of a protein, suitable for mutating to more hydrophilic residues (e.g., charged amino acids). Any number of residues on the surface may be changed to create a variant that is more soluble, resistant to aggregation, has a greater ability to re-fold, and/or is more stable under a variety of conditions. The invention also provides GFP, streptavidin, and GST variants with an increased theoretical net charge created by the inventive technology. Kits are also provided for carrying out such modifications on any protein of interest.

Claims35

Dependent claims

A method of improving the stability of a protein of interest, the method comprising steps of:

(1) identifying non-conserved surface residues of a protein of interest by comparing the amino acid sequence of the protein with at least one other amino acid sequence of the protein from the same protein family or a different species, wherein a residue is non-conserved if less than or equal to 50% of the amino acid sequences have the same amino acid sequence in a particular position;
and
(2) replacing a plurality of non-conserved, surface residues identified in step (1) with a natural amino acid residue that is positively charged at physiological pH, whereby the method creates a modified protein of interest having a theoretical net charge of at least about +10 at physiological pH as compared to the theoretical net charge of the original protein of interest.

Dependencies are inferred from the claim text.

Protein surface remodeling

Life of this patent

Protein surface remodeling

Life of this patent

Abstract

Claims35