Emd Millipore Corporation

Millipore Corporation

Upfront Chromatography A/s

Neil P Soice

Nanying Bian

Wilson Moya

John Charkoudian

Chen Wang

Joaquin A Umana

Kwok-Shun Cheng

Volkmar Thom

Senthilkumar Ramaswamy

Brian Gagnon

Anthony J DiLeo

James Hamzik

Justin McCue

Igor Quinones-Garcia

Sarah Yuan

Yu Zhang

Jad Jaber

Nripen Singh

Dennis Aquino

Arezki Boudif

Philip M Goddard

Shari Spector

Andrew Lyddiatt

Antoni Peters

Dave Brewster

Yuan Wong

Kara Levine

Anja Dedeo

Senthil Ramaswamy

John Dana Hubbard

Umana Joaquin

Lyddiatt Andrew

Senthikumar Ramaswamy

The present invention relates to chromatography ligands having improved caustic stability, e.g., ligands based on immunoglobulin-binding proteins such as, Staphylococcal protein A, as well as methods of making and using such ligands.

Caustic stable chromatography ligands

The present invention relates to chromatography ligands having improved caustic stability, e.g., ligands based on immunoglobulin-binding proteins such as,protein A, as well as methods of making and using such ligands.

The present invention provides novel and improved stimulus responsive polymers and methods of using the same for the purification of biomolecules.

Stimulus responsive polymers for the purification of biomolecules

The invention relates generally to solid supports for chromatography. In specific embodiments the invention provides for solid supports suitable for affinity chromatography along with methods, systems and kits which use the same.

Media for affinity chromatography

A porous substrate capable of adsorptive filtration of a fluid having a porous self-supporting substrate and one or more porous, adsorptive polymeric coatings comprising from about 1 to about 80% of the void volume of the pores of the substrate. The resultant substrate has good convective and diffusive flow and capacity. The substrate may be cross-linked, have one or more capture chemistries attached to it and is useful as a chromatography media for the selective filtration of desired species including biomolecules such as proteins and DNA fragments.

Porous adsorptive or chromatographic media

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

A method for making functionalized porous crosslinked polysaccharide gel coated structures used as liquid chromatography media is provided. The method includes impregnating a porous substrate with a room temperature stable aqueous polysaccharide solution containing water, 0.1% to 20% of a polysaccharide, 18% to 54% of a gel-inhibiting agent to prevent the gel from re-gelling, and 0.001% to 10% of an anionic fluorosurfactant for optimum solution coatability onto the substrate, each concentration is by total weight of the aqueous solution. Next water is evaporated from the coating, followed by exposing the dehydrated coating to a gelling agent thereby forming a porous polysaccharide gel coated substrate. Next the gel coated substrate is exposed to a crosslinking agent forming a porous crosslinked polysaccharide gel coated substrate. Next, the gel coated substrate is functionalized by contacting the porous polysaccharide gel coated substrate with sodium 3-bromopropanesulfonate, thereby attaching sulfopropyl ligands to the gel coated substrate, resulting in a functionalized porous absorptive polysaccharide gel coated chromatography structure. When the functionalized gel coated chromatography structure is used as liquid chromatography media, and a protein (e.g. lysozyme) containing solution contacts the media, the sulfopropyl ligands attached to the gel coated chromatography structure will bond to the protein, thereby removing the protein from the protein containg solution.

Room temperature stable agarose solutions

The present invention relates, at least in part, to improved methods of protein purification. In particular, the present invention relates, at least in part, to methods for purifying an Fc region containing protein from a composition comprising the Fc region containing protein and one or more impurities, where the methods eliminate the need for a holding tank and/or a buffer exchange step.

Methods for purifying a target protein from one or more impurities in a sample

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

Hydrophilic membrane particularly suited for blotting applications, preferably Western blotting. A pre-wet hydrophobic membrane substrate, preferably made of PVDF, is contacted with a monomer solution and subjected to a UV-initiated free radical polymerization step to render the substrate permanently hydrophilic. The resulting membrane exhibits low background fluorescence, high protein binding, excellent retention of protein sample spot morphology, and extended dynamic range (high signal-to-noise ratio, enhanced sample detectability). The membrane demonstrates comparable or higher performance in Western blotting applications than conventional nitrocellulose Western blotting membranes, particularly for protein detection at low sample concentrations, and is directly water-wettable, eliminating the need for an alcohol pre-wet step prior to use.

Hydrophilic, high protein binding, low fluorescence, western blotting membrane

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Merrimack, NH, United States of America

Neil P Soice