Genencor International, Inc.

Dow Corning Corporation

Danisco US Inc.

Meng Hong Heng

Anthony Revis

Thomas Howard Lane

Csilla Kollar

Gary L Gibson

Keith Hayes

Landon M Steele

Nathaniel T Becker

Richard Ray Bott

Christian G Paech

Robert I Christensen

Robin Fong

Edit L Braunstein

Antti V Kosola

Robert C Fewkes

Ernst H Gros

Shauna L Bowden

Michael Bodo

Kevin Dibel

Glenn Rozeboom

The present methods relate to the isolation and concentration of proteins using cross-flow membrane filtration, and to proteins produced by such methods. A feature of the method is that the protein of interest is retained by a cross-flow membrane under certain conditions that promote retention, while under other condition the protein passes through the membrane.

Cross-flow membrane filtration-based process for protein recovery

The present invention provides methods for separating one or more components of interest from a sample containing particulates and soluble materials. The method comprises the steps of: (a) filtering a sample through silica filter media whose surface silanol groups have reacted with one or more silanes, and (b) simultaneously capturing particulates and binding a soluble component to the silica filter media. The bound soluble component of interest is subsequently eluted from the silica filter media. In one embodiment of the invention, unwanted soluble materials are captured by the treated silica filter media and desired component of interest is recovered from the flow-through. In another embodiment, different components of interest are recovered from both the eluate and the flow-through. Preferred treated silica filter media are silane-treated rice hull ash or diatomaceous earth with functional quarternary ammonium group or functional sulphonate group. The present invention also provides silane-treated silica filter media.

Silane-treated silica filter media

The present invention relates to a method of purifying a protein of interest from its fusion analog, comprising obtaining a protein solution comprising a protein of interest and its fusion analog; adjusting the pH and/or ionic strength of the protein solution with an appropriate buffer for use with a Hydrophobic Charge Induction Chromatography (HCIC) resin, contacting the protein solution and HCIC resin column to allow binding of the protein of interest and its fusion analog, washing the resin and eluting the protein of interest from the resin by a pH gradient; wherein the protein of interest is substantially free of its fusion analog.

Protein purification

The present invention provides methods for separating one or more components of interest from a sample containing particulates and soluble materials. The method comprises the steps of: (a) filtering a sample through silica filter media whose surface silanol groups have reacted with one or more silanes, and (b) simultaneously capturing particulates and binding a soluble component to the silica filter media. The bound soluble component of interest is subsequently eluted from the silica filter media. In one embodiment of the invention, unwanted soluble materials are captured by the treated silica filter media and desired component of interest is recovered from the flow-through. In another embodiment, different components of interest are recovered from both the eluate and the flow-through. Preferred treated silica filter media are silane-treated rice hull ash or diatomaceous earth with functional quarternary ammonium group or functional sulphonate group. Particulates suitable for the present invention, for example, are microorganisms.

Method of separating components in a sample using silane-treated silica filter media

The present invention provides methods for separating one or more components of interest from a sample containing particulates and soluble materials. The method comprises the steps of: (a) filtering a sample through silica filter media whose surface silanol groups have reacted with one or more silanes, and (b) simultaneously capturing particulates and binding a soluble component to the silica filter media. The bound soluble component of interest is subsequently eluted from the silica filter media. In one embodiment of the invention, unwanted soluble materials are captured by the treated silica filter media and desired component of interest is recovered from the flow-through. In another embodiment of the invention, different components of interest are recovered from both the eluate and the flow-through. Preferred treated silica filter media are silane-treated rice hull ash or diatomaceous earth with functional quaternary ammonium group or functional sulphonate group. Particulates suitable for the present invention, for example, are microorganisms.

The present invention relates to a method of purifying a protein of interest from its fusion analog, comprising obtaining a protein solution comprising a protein of interest and its fusion analog; adjusting the pH and/or ionic strength of the protein solution with an appropriate buffer for use with a Hydrophobic Charge Induction Chromatograph (HCIC) resin, contacting the protein solution and HCIC resin column to allow binding of the protein of interest and its fusion analog, washing the resin and eluting the protein of interest from the resin by a pH gradient; wherein the protein of interest is substantially free of its fusion analog.

Enzyme-containing formulations having improved stability and enzymatic activity in liquid medium, comprising one or more protease enzymes produced from anybacteria, at least about 5% alkali metal halide salt, and at least about 50% polyol. Also disclosed are methods for making such formulations.

Stabilization of enzymes

The present invention provides a crystallization process wherein a starting temperature is selected such that a desirable crystal morphology (e.g., square) is obtained. A temperature shift is then introduced, providing that the shift is not enough to induce further nucleation, where the crystals continue to grow in the desirable fashion, but with different kinetics, e.g., a higher rate of crystallization. As a result, the process gives a crystalline product with desirable morphology at a higher crystallization rate.

Method for rapidly obtaining crystals with desirable morphologies

The present invention provides a crystallization process wherein a starting temperature is selected such that a desirable crystal morphology (e.g., square) is obtained. A temperature shift is then introduced, providing that the shift is not enough to induce further nucleation, where the crystals continue to grow in the desirable fashion, but with different kinetics, e.g., a higher rate of crystallization. As a result, the process gives a crystalline product with desirable morphology at a higher crystallization rate. The starting temperature of the process can be between about 4&deg; C. and 20&deg; C. for no more than about 5 hours and the temperature shift of the process can be between about 22&deg; C. and 60&deg; C. for no more than about 20 hours.

Method for rapidly obtaining enzyme crystals with desirable morphologies

A method for preparing a crystalline cellulase enzyme is provided which comprises preparing an aqueous solution containing cellulase enzyme and adding to the aqueous solution a salt comprising an anion selected from the group consisting of sulfate, phosphate, formate, acetate, sorbate, chloride, bromide, fluoride or iodide, and a cation selected from the group consisting of sodium, ammonium, magnesium, potassium or calcium, wherein the aqueous solution is at a temperature between 10&deg; C. and 60&deg; C.

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Belmont, CA, United States of America

Meng Hong Heng