Emd Millipore Corporation

Millipore Corporation

Upfront Chromatography A/s

Brian Gagnon

Senthilkumar Ramaswamy

Nanying Bian

Neil P Soice

Kwok-Shun Cheng

Joaquin A Umana

Wilson Moya

Mikhail Kozlov

Chen Wang

Michael W Phillips

Dennis Aquino

Andrew Lyddiatt

Umana Joaquin

Lyddiatt Andrew

Senthikumar Ramaswamy

Media and devices, such as anion exchangers including such media, wherein the media is a membrane having a surface coated with a polymer such as a polyallylamine. The resulting membrane offers stronger binding of protein impurities and superior removal of host cell proteins from biological samples than conventional ligands based on quaternary ammonium salts, including trimethylammonium ligands. Also described is a chromatography scheme and method for purifying monoclonal antibodies, wherein the anion exchange sorber is placed downstream of an affinity column (such as Protein A or Protein G affinity column) and optionally one or more polishing devices such as cationic exchange columns. Little or no dilution of the cation exchanger pool (or affinity column exchange pool where no cation exchanger is used) is necessary to lower the conductivity of the sample. The sorber functions well to strongly bind host cell proteins and other impurities in biological samples even at high conductivities and pH.

Media for membrane ion exchange chromatography based on polymeric primary amines, sorption device containing that media, and chromatography scheme and purification method using the same

The present invention relates to an asymmetric chromatography media suitable for separations applications, particularly as packed bed, fluidized bed or magnetized bed chromatography media. In certain embodiments, the asymmetric chromatography media comprises asymmetric particles, preferably beads, having at least two distinct, controlled pore size distributions. Preferably one of the distinct pore size distributions is in an internal region of the particle, and the other is in an external region or coating on the particle. These distinct pore size distributions can be modified with uniform or alternatively unique functional groups or mixtures of functional groups. The present invention allows for the control over pore size distribution within an asymmetric porous particle by providing a distinct internal region, preferably in the form of a bead, and a distinct external region, preferably in the form of a coating on the bead.

Asymmetric porous adsorptive bead

Media and devices, such as anion exchangers including such media, wherein the media is a membrane having a surface coated with a polymer such as a polyallylamine. The resulting membrane offers stronger binding of protein impurities and superior removal of host cell proteins from biological samples than conventional ligands based on quaternary ammonium salts, including trimethylammonium ligands. Also described is a chromatography scheme and method for purifying monoclonal antibodies, wherein the anion exchange sorber is placed downstream of an affinity column (such as Protein A or Protein G affinity column) and optionally one or more polishing devices such as cationic exchange columns. Little or no dilution of the cation exchanger pool (or affinity column exchange pool where no cation exchanger is used) is necessary to lower the conductivity of the sample. The sorber functions well to strongly bind host cell proteins and other impurities even at high conductivities and pH.

The present invention relates to a method and apparatus for forming agarose or cored agarose beads. The process involves dissolving/gelation the agarose in a suitable liquid, mixing it with a hydrophobic liquid to form an emulsion and maintaining that emulsion at a temperature equal to or greater than the gelation point of the agarose, passing it through a static mixer to create agarose droplets and solidifying the agarose droplets in a second bath of hydrophobic liquid. The beads can then be washed and used or further processed to crosslink the agarose and/or add various functionalities on to the agarose. Another method for solidifying the agarose droplets is by using a heat exchanger to cool the stream continuously after it exits the static mixer. A similar process is used for the 'cored' beads except cores, preferably in bead form, are introduced to the agarose before it enters the first hydrophobic liquid so that the agarose forms a coating on the cores. A similar process with either agarose beads (made by this or another process) or cored agarose (made by this or another process) can be used to add multiple layers of agarose on to the existing beads. An apparatus for running the process is also disclosed.

Method and apparatus for making porous agarose beads

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Billerica, MA, United States of America

Brian Gagnon