Compact hydroxamate-based affinity tags for artificially tagging biological macromolecules
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsBjorn-Erik Wulff, Jenna G. Caldwell, Pehr B. Harbury
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Abstract
Methods for purifying biological macromolecules are provided. Aspects of the subject methods include contacting the biological macromolecule with an exemplary hydroxamate affinity tag to produce a tagged moiety followed by purification of the tagged moiety by immobilized metal affinity chromatography (IMAC). Also provided are kits comprising an exemplary subject hydroxamate affinity tag, an IMAC resin and a metal ion configured for loading onto the resin, wherein the metal ion is capable of binding to a compound containing the hydroxamate affinity tag.
Claims20
A method of purifying a biological macromolecule, the method comprising:
- (a) contacting the biological macromolecule with a compound of formula (I):
- wherein:
- Y is a selective reactive group configured to react with the biological macromolecule chemically or chemo-enzymatically to produce a tagged moiety;
- R is selected from hydrogen, alkyl or substituted alkyl;
- and
- (b) purifying the tagged moiety by immobilized metal affinity chromatography (IMAC).
A kit comprising:
- a compound of formula (I)
- wherein:
- Y is a selective reactive group configured to react with a biological macromolecule chemically or chemo-enzymatically;
- R is selected from hydrogen, alkyl and substituted alkyl;
- and
- an immobilized metal affinity chromatography (IMAC) resin;
- and
- a metal ion configured for loading onto the resin, wherein the metal ion is capable of binding a compound of formula (I).
Dependencies are inferred from the claim text.