US 12,472,448 B2
Compact hydroxamate-based affinity tags for artificially tagging biological macromolecules
Bjorn-Erik Wulff, East Palo Alto, CA (US); Jenna G. Caldwell, Bethesda, MD (US); and Pehr B. Harbury, Portola Valley, CA (US)
Assigned to THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY, Stanford, CA (US)
Appl. No. 17/413,912
Filed by The Board of Trustees of the Leland Stanford Junior University, Stanford, CA (US)
PCT Filed Jan. 23, 2020, PCT No. PCT/US2020/014847
§ 371(c)(1), (2) Date Jun. 14, 2021,
PCT Pub. No. WO2020/154541, PCT Pub. Date Jul. 30, 2020.
Claims priority of provisional application 62/796,430, filed on Jan. 24, 2019.
Claims priority of provisional application 62/796,403, filed on Jan. 24, 2019.
Claims priority of provisional application 62/796,475, filed on Jan. 24, 2019.
Claims priority of provisional application 62/796,424, filed on Jan. 24, 2019.
Prior Publication US 2022/0054955 A1, Feb. 24, 2022
Int. Cl. B01D 15/38 (2006.01); B01J 20/02 (2006.01); B01J 20/281 (2006.01); C07K 1/22 (2006.01)
CPC B01D 15/3828 (2013.01) [B01J 20/0207 (2013.01); B01J 20/0225 (2013.01); B01J 20/281 (2013.01); C07K 1/22 (2013.01); B01J 2220/4806 (2013.01)] 12 Claims
 
1. A method of purifying a biological macromolecule, the method comprising:
(a) contacting the biological macromolecule with a compound of formula (II) or formula (III):

OG Complex Work Unit Chemistry
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;

OG Complex Work Unit Chemistry
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).