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.
Claims12
A method of purifying a biological macromolecule, the method comprising:
- (a) contacting the biological macromolecule with a compound of formula (II) or formula (III):
- wherein:
- L is an optional linker,
- R1 is an optional group selected from halogen, deuterium, alkyl and substituted alkyl, and
- n is an integer from 0 to 3;
- wherein:
- T is an optional linker/tether,
- L is a leaving group,
- R1 is an optional group selected from halogen, deuterium, alkyl and substituted alkyl, and
- n is an integer from 0 to 8;
- to produce a tagged moiety that comprises the biological macromolecule and a hydroxamate group;
- and
- (b) purifying the tagged moiety by immobilized metal affinity chromatography (IMAC).
A kit comprising:
- a compound of formula (II) or formula (III):
- wherein:
- L is an optional linker,
- R1 is an optional group selected from halogen, deuterium, alkyl and substituted alkyl, and
- n is an integer from 0 to 3;
- wherein:
- T is an optional linker/tether,
- L is a leaving group,
- R1 is an optional group selected from halogen, deuterium, alkyl and substituted alkyl, and
- n is an integer from 0 to 8;
- 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 (II) or (III).
Dependencies are inferred from the claim text.