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Use of small molecules in methods for purification of biomolecules

A technology of biomolecules and small molecules, applied in the field of the use of small molecules in the purification of biomolecules, can solve the problems of difficult removal, expensive polymers, and toxic, and achieve the effect of easy operation

Inactive Publication Date: 2014-04-16
MILLIPORE CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The main drawback of this flocculation technique is that it requires the use of polymers that can become entangled with the target protein, be toxic and / or not easily cleared from the patient's body, which are expensive in single-use applications , and is not readily available as it usually requires the synthesis of

Method used

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  • Use of small molecules in methods for purification of biomolecules
  • Use of small molecules in methods for purification of biomolecules
  • Use of small molecules in methods for purification of biomolecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] Example 1: Preparation of expression cell culture fluid (CCF)

[0124] In a representative experiment, express monoclonal IgG 1 Of cells derived from the Chinese Hamster Ovary (CHO) cell line were grown to 13×10 in a 10L bioreactor (NEW BRUNSWICK SCIENTIFIC) 6 The density of cells / ml and in Harvested at <50% survival rate. The antibody titer was determined by protein AHPLC in the range of 0.85-1.8mg / mL. Using ELISA (CYGNUS#F550) found that the level of host cell protein (HCP) was 350000-425000ng / mL. The pH of the unclarified cell culture medium was pH 7.2.

Embodiment 2

[0125] Example 2: Preparation of Clarified Cell Culture Medium Fluid (CCF)

[0126] By centrifuging at 4000 rpm for 2 min, then passing 5 μm and 0.2 μm The filter was filtered to clarify the feed of Example 1.

Embodiment 3

[0127] Example 3: Preparation of unexpressed cell culture fluid (CCF)

[0128] In another experiment, cells derived from a non-IgG expressing Chinese Hamster Ovary (CHO) cell line were grown to 13×10 in a 10L bioreactor (NEW BRUNSWICK SCIENTIFIC) 6 The density of cells / ml and in Harvested at <50% survival rate. Using ELISA (CYGNUS#F550) found that the level of host cell protein (HCP) was 66000-177000ng / mL. The pH of the unclarified cell culture medium was pH 7.2.

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Abstract

The present invention relates to novel and improved methods for the purification of biomolecules. In particular, the present invention relates to methods of protein purification which employ small molecules, which include at least one non-polar group and at least one cationic group or which include at least one non-polar group and at least one anionic group.

Description

[0001] Related application [0002] This patent application claims the priority of U.S. Provisional Patent Application No. 61 / 575,376 filed on August 19, 2011 and U.S. Provisional Patent Application No. 61 / 666,287 filed on June 29, 2012, the entire contents of which are hereby quoted Join. Technical field [0003] The present invention relates to a novel and improved method of purifying biomolecules. In particular, the present invention relates to methods of purifying proteins with small molecules. Background technique [0004] The general method of manufacturing biomolecules such as proteins and especially recombinant therapeutic proteins typically includes two main steps: (1) expression of the protein in the host cell, and (2) purification of the protein. The first step usually involves growing the desired host cells in a bioreactor to facilitate the expression of the protein of interest. Once the protein is expressed at the desired level, the protein is removed from the host c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P35/00
CPCA61K39/395C07K16/00B01D15/327B01D15/361B01D15/3804B01D15/3847C07K1/30A61P35/00
Inventor J·贾比尔W·莫亚A·波蒂A·杜邦M·T·斯通M·科兹洛夫
Owner MILLIPORE CORP