US 12,134,089 B2
Multimodal chromatographic media for protein separation
Christopher A. Pohl, Union City, CA (US); Xiaodong Liu, Cupertino, CA (US); and Shanhua Lin Liehr, Palo Alto, CA (US)
Appl. No. 16/477,752
Filed by DIONEX CORPORATION, Sunnyvale, CA (US)
PCT Filed Jan. 18, 2018, PCT No. PCT/US2018/014284
§ 371(c)(1), (2) Date Jul. 12, 2019,
PCT Pub. No. WO2018/136666, PCT Pub. Date Jul. 26, 2018.
Claims priority of provisional application 62/448,397, filed on Jan. 20, 2017.
Prior Publication US 2019/0358621 A1, Nov. 28, 2019
Int. Cl. B01J 39/20 (2006.01); B01D 15/36 (2006.01); B01D 15/38 (2006.01); B01J 20/26 (2006.01); B01J 20/28 (2006.01); B01J 20/288 (2006.01); B01J 20/32 (2006.01); B01J 39/26 (2006.01); B01J 41/14 (2006.01); B01J 41/20 (2006.01); C07K 1/18 (2006.01)
CPC B01J 39/20 (2013.01) [B01D 15/362 (2013.01); B01D 15/363 (2013.01); B01D 15/3847 (2013.01); B01J 20/261 (2013.01); B01J 20/28019 (2013.01); B01J 20/288 (2013.01); B01J 20/321 (2013.01); B01J 20/3217 (2013.01); B01J 20/3278 (2013.01); B01J 20/3285 (2013.01); B01J 20/3289 (2013.01); B01J 20/3293 (2013.01); B01J 20/3295 (2013.01); B01J 39/26 (2013.01); B01J 41/14 (2013.01); B01J 41/20 (2013.01); C07K 1/18 (2013.01); B01J 2220/445 (2013.01)] 24 Claims
OG exemplary drawing
 
1. A chromatographic media for separating bio-polymers, the chromatographic media having cationic and anionic retention modes, the chromatographic media comprising:
non-porous substrate particles including an organic polymer, the non-porous substrate particles having a neutral hydrophilic layer at an external surface thereof, the neutral hydrophilic layer configured to reduce a binding of the bio-polymers directly to the non-porous substrate particles compared to a binding of the bio-polymers to the non-porous substrate particles without the neutral hydrophilic layer;
a charged first ion exchange layer bound to the substrate particles on top of the neutral hydrophilic layer, the first ion exchange layer comprising a plurality of polymer chains, each polymer chain of the plurality of polymer chains having a length and being bound to the substrate particles such that each of the polymer chains extends away from the substrate particles in a direction orthogonal to the external surfaces thereof, and each polymer chain comprising between 5 and 100 first ion exchange groups distributed along the length thereof;
a charged second ion exchange layer comprising charged nano-particles bound to the substrate particles on top of the first ion exchange layer, the second ion exchange layer comprising second ion exchange groups, the second ion exchange layer having an opposite charge to the first ion exchange layer and being bound to the substrate particles by electrostatic interactions with the first ion exchange layer,
wherein the first ion exchange layer provides one of the cationic and anionic retention modes and the second ion exchange layer provides the other one of the cationic and anionic retention modes,
wherein the first and second ion exchange layers both reside on the exterior surface of the non-porous substrate particles and are configured such that for each charged nano-particle of the second ion exchange layer, the charged nano-particle partially interpenetrates a respective group of polymer chains of the plurality of polymer chains of the first ion exchange layer and electrostatically interacts with a first portion of the between 5 and 100 of the first ion exchange groups carried by each of the polymer chain of said respective group of polymer chains, and such that a second portion of the between 5 and 100 of the first ion exchange groups carried by each of the polymer chains of the same respective group of polymer chains remain spatially separated from the charged nano-particles, such that in aggregate the charged nano-particles of the second ion exchange layer reduce the retention properties of the first ion exchange layer, as measured by retention time, by less than 50%,
wherein the respective group of polymer chains of at least some different charged nano-particles are non-overlapping one with another,
and wherein both the cationic and anionic retention modes are accessible to bind the bio-polymers.