Hoffmann-la Roche Inc.

Genentech, Inc.

F. Hoffmann-la Roche Ag

Maxygen, Inc.

Roberto Falkenstein

Markus Strasser

Phillip A Patten

Stefan Seeber

Stephan Fischer

Tilman Schlothauer

Ralf Schumacher

Petra Rueger

Thomas Meier

Adelbert Grossmann

Torben Lauesgaard Nissen

Rainer Schmuck

Wilhelm Tischer

Claudia Giessel

Bernhard Spensberger

Thorsten Lemm

Klaus Schwendner

Christian Schantz

Andreas Schaubmar

Sridhar Viswanathan

Friederike Hesse

Hartmut Duefel

Michael Wiedmann

Hidenari Yamada

Martin Bader

Marco Thomann

Sebastian Malik

Andreas Blaschyk

Nicole Fuehrler

Wolfgang Kuhne

Hartmut Ernst Schurig

Anne Vogt

Harald Kropshofer

Hubert Hertenberger

Benjamin Tran

Thomas Dams

Wolfgang Koehnlein

Birgit Weydanz

Marc Pompiati

Marc Wong

Maria Sebald

Burkard Kolb

Heiko Walch

Ingrid Grunert

Matthias Freiherr Von Roman

Michael Leiss

Christian Meyer

Alexandra Thomas Morris

Michaela Mehr

Jie Gu

Jeevan Prabhakar

Yinges Yigzaw

Franziska Edelmann

Tobias Graf

Maria Smida

Brigitte Kraemer

Sybille Gabel

Maria Laura Magri

Iris Lein

Hartmut Schurig

Blaschyk Andreas

Ryan Erickson

Paul Mcdonald

Richard St John

Tarik Khan

Sanjay Gupta

Sybille Greithanner

Hartmut (deceased) Schurig

Roland Dorn

Michael Hingar

Carina Kopp

Annika Kleinjans

Frank Zettl

Lars Hillringhaus

The present disclosure provides purification platforms comprising a depth filter step and/or a hydrophobic interaction chromatography (HIC) step. Also disclosed herein are methods of using the purification platforms described herein and compositions obtained therefrom, such as pharmaceutical compositions.

Methods of reducing the enzymatic hydrolysis activity rate in a composition obtained from a purification platform

Herein is reported a method for the purification of an antibody directly captured from clarified cell culture supernatants using Streamline CST and/or Capto MMC, wherein especially product related (aggregates and fragments) and process related impurities (host cell protein, media components) could efficiently be removed, resulting in a preparation with a purity comparable to classical protein A affinity chromatography.

Optimized method for antibody capturing by mixed mode chromatography

Herein is reported a method for the enzymatic preparation/production of an antibody with a modified glycosylation in the Fc-region comprises the steps of incubating an antibody that has a glycosylation in the Fc-region with one or more enzymes for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating in one or more chromatography steps the antibody with a modified glycosylation in the Fc-region from the one or more enzymes and thereby producing the antibody with a modified glycosylation in the Fc-region and one or more recycled enzymes, and repeating the incubating step with the one or more recycled enzymes at least once.

Re-use of enzymes in in vitro glycoengineering of antibodies

Herein is reported a method for the purification of a protein comprising erythropoietin and a single poly (ethylene glycol) residue from reaction by-products or not reacted starting material by a cation exchange chromatography method. It has been found that by employing a cation exchange Toyopearl® SP-650 chromatography material and employing a second wash step with an increased pH value compared to the first wash step a fusion protein of erythropoietin and a single poly (ethylene glycol) residue can be obtained in a single step with high purity and yield and suitability for large scale applications.

Method for purifying PEGylated erythropoietin

Herein is reported a method for the final filtration of concentrated polypeptide solutions comprising the combination of two immediately consecutive filtration steps with a first filter of 3.0 μm and 0.8 μm pore size and a second filter of 0.45 μm and 0.22 μm pore size.

Multistep final filtration

Herein is reported the use of an immobilized non-covalent complex of a neonatal Fc receptor (FcRn) and beta-2-microglobulin (b2m) as affinity chromatography ligand in general and, for example, for the determination of the in vivo half-live of an antibody by determining the ratio of the retention times of the antibody and a reference antibody.

Fc-receptor based affinity chromatography

Herein is reported a method for the enzymatic production of an antibody with a modified glycosylation in the Fc-region comprising the steps of incubating the antibody light chain affinity ligand-bound monoclonal antibody with a glycosylation in the Fc-region with a first enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region, recovering the antibody from the antibody light chain affinity ligand, incubating the recovered antibody in solution with a second enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating the antibody with the modified glycosylation in the Fc-region from the second enzyme in a cation exchange chromatography, and thereby producing the antibody with a modified glycosylation in the Fc-region.

Method for in vitro glycoengineering of antibodies

Herein is reported a method for purifying a polypeptide comprising the steps of i) applying a solution comprising the polypeptide to an ion exchange chromatography material, and ii) recovering the polypeptide with a solution comprising a denaturant and thereby purifying the polypeptide, whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached, and wherein the solution comprising the polypeptide applied to the ion exchange chromatography material is free of the denaturant and the polypeptide adsorbed to the ion exchange chromatography material is recovered with a solution comprising a denaturant at a constant conductivity.

Cation and anion exchange chromatography method

The current invention reports a method for purifying an immunoglobulin, wherein the method comprises applying an aqueous, buffered solution comprising an immunoglobulin in monomeric and in aggregated form to a cation exchange material under conditions whereby the immunoglobulin in monomeric form does not bind to the cation exchange material, and recovering the immunoglobulin in monomeric form from the solution after the contact with the cation exchange material.

Immunoglobulin purification

A method for the purification of immunoglobulins by ion exchange chromatography is described. The chromatographic method uses a weak ion exchange resin and a single step elution process for the purification of an immunoglobulin. Additionally a method for the determination of the salt concentration for the single step elution of an immunoglobulin from an ion exchange resin is described.

Method for the purification of antibodies

Herein is reported a method for the purification of a protein comprising erythropoietin and a single poly (ethylene glycol) residue from reaction by-products or not reacted starting material by a cation exchange chromatography method. It has been found that by employing a cation exchange SP Sephacryl&#x2122; S 500 HR chromatography material conditioned to a conductivity of 21 mS/cm and a linear gradient elution a fusion protein of erythropoietin and a single poly (ethylene glycol) residue can be obtained in a single step with high purity and yield.

Herein is reported a method for producing an antibody preparation comprising the steps of a) applying a buffered solution comprising different isoforms of an antibody to a cation exchange chromatography material, b) applying a first solution with a first conductivity to the cation exchange chromatography material, whereby the antibody isoforms remain bound to the cation exchange chromatography material, and c) applying a second solution with a second conductivity to the cation exchange chromatography material and thereby obtaining the antibody preparation, whereby the conductivity of the second solution exceeds the conductivity of the first solution by not more than 10%.

Isoform enriched antibody preparation and method for obtaining it

The current invention is directed to a chromatography column separator which separates the chromatography column in an upper chromatography column chamber and a lower chromatography column chamber, and which has a variable position within the chromatography column, and which is embedded by the chromatography material. The separator allows the replacement of the chromatography material in the upper chromatography column chamber without the need to replace the chromatography column material in the lower chromatography column chamber and it allows also the combination of two different chromatography materials with different chromatographical functional groups in one chromatography column.

Moveable chromatography column separator

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Munich, Germany

Roberto Falkenstein