US 11,857,619 B2
Multimodal chromatography method for the purification of HIV-1 envelope glycoprotein
Christopher Rode, Downingtown, PA (US); Brian Polilli, Phoenixville, PA (US); and John Schreffler, Downingtown, PA (US)
Assigned to Janssen Vaccines & Prevention B.V., Leiden (NL)
Filed by Janssen Vaccines & Prevention B.V., Leiden (NL)
Filed on Nov. 6, 2020, as Appl. No. 17/091,296.
Claims priority of provisional application 63/105,664, filed on Oct. 26, 2020.
Claims priority of provisional application 62/932,180, filed on Nov. 7, 2019.
Prior Publication US 2021/0138060 A1, May 13, 2021
Int. Cl. A61K 39/21 (2006.01); C07K 14/16 (2006.01); C12N 7/02 (2006.01); C07K 1/18 (2006.01); C07K 14/005 (2006.01); A61K 39/00 (2006.01)
CPC A61K 39/21 (2013.01) [C07K 1/18 (2013.01); C07K 14/005 (2013.01); C07K 14/162 (2013.01); C12N 7/02 (2013.01); A61K 2039/53 (2013.01); C12N 2740/16051 (2013.01); C12N 2740/16111 (2013.01); C12N 2740/16122 (2013.01); C12N 2740/16134 (2013.01)] 14 Claims
 
1. A process for purifying HIV-1 gp140 protein, comprising the steps of:
i) providing a composition comprising HIV-1 gp140 protein and other, non-desired host cell proteins derived from the host cell in which HIV-1 gp140 protein is expressed;
ii) capturing the HIV-1 gp140 protein on a multimodal resin comprising hydrophobic interaction and cation exchange properties, and eluting a purified fraction comprising the HIV-1 gp140 protein from the multimodal resin, wherein the multimodal resin comprising hydrophobic interaction and cation exchange properties comprises a solid support matrix with the following attached ligand of

OG Complex Work Unit Chemistry
iii) applying the purified fraction of step ii) to an anion exchange resin to bind the HIV-1 gp140 protein, and eluting a further purified fraction comprising the HIV-1 gp140 protein from the anion exchange resin, wherein the anion exchange resin comprises a solid support matrix with a ligand of quaternary polyethyleneimine;
iv) applying the further purified fraction of HIV-1 gp140 protein of step iii) to an affinity medium resin comprising the ligand dextran sulfate, and eluting a further purified fraction comprising the HIV-1 gp140 protein from said resin; and,
v) subjecting the further purified fraction of step iii) to a multimodal resin that has anion-exchange and hydrophobic functionalities, wherein the multimodal resin that has anion-exchange and hydrophobic functionalities comprises a solid support matrix with a ligand of

OG Complex Work Unit Chemistry
and eluting a further purified HIV-1 gp140 protein.