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Removal of virucidal agents in mixed mode chromatography

A virucidal, hybrid technology, applied in peptides, immunoglobulins, chemical instruments and methods, etc., can solve the problems of difficulty and impossibility of removing virucidal agents

Inactive Publication Date: 2014-07-02
BIO RAD LAB INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These combinations may make complete removal of virucides from protein formulations difficult or impossible

Method used

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  • Removal of virucidal agents in mixed mode chromatography
  • Removal of virucidal agents in mixed mode chromatography

Examples

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Embodiment 1

[0096] Example 1: Removal of Virucides from Antibody Preparations

[0097] This example describes the use of a mixed support to remove virucidal agents from antibody formulations.

[0098] Biomolecule preparations including IgM antibodies were treated with the viricide PEI-1300 (average molecular weight 1300 Daltons) at a concentration of 0.01%. Equilibrium with 20 mM MES, 20 mM acetate includes the macromolecules produced by the polymerization of the monomers 3-allyloxy-1,2-propanediol, vinylpyrrolidone and cross-linked with N,N'-methylenebisacrylamide. Porous matrix-linked ligand p-aminohippuric acid column to pH 5.0. The pH of the sample was lowered to pH 4.75 by the addition of 1 M acetate, pH 4.5, 5% vol:vol, and the sample was loaded onto a mixed support column. An intermediate 1M NaCl wash was applied. IgM antibodies were eluted by increasing the operating pH to 7.5 in a linear gradient of 20 mM Hepes. Antibody purity was estimated to be greater than 90% by analytic...

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Abstract

The present invention provides chromatography methods for removing a virucidal agent from a target protein such as an antibody. The method uses a hydrophobic, negatively charged mixed mode support that has high affinity for both the target protein and the virucidal agent. The method provides conditions that favor dissociation of the virucidal agent from the protein, allowing the virucidal agent to bind strongly to the support. The target protein is then eluted from the support under conditions such that the virucidal agent remains bound to the support.

Description

[0001] Cross References to Related Patent Applications [0002] This application claims priority to US Provisional Patent Application No. 61 / 551.735, filed October 26, 2011, which is incorporated herein by reference for all purposes. technical background [0003] Native and recombinant proteins produced by in vivo or in vitro methods require treatment with virucidal conditions or compounds to ensure the safety of patients receiving treatments based on these proteins. Many viricides are toxic. A complication of virucidal therapy is that the virucidal agent itself may form stable associations with the protein product of the treatment. These combinations may make it difficult or impossible to completely remove the virucide from the protein formulation. Contents of the invention [0004] Methods for removing cationic or neutral virucides from biomolecular formulations are provided. [0005] In some embodiments, the method comprises contacting a biomolecule preparation compris...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00
CPCC07K16/065
Inventor P·S·加农
Owner BIO RAD LAB INC