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Methods of measuring hydrophobicity of chromatographic resin

A technique of hydrophobicity and chromatography, applied in the field of measuring the hydrophobicity of chromatographic resins

Pending Publication Date: 2021-11-12
BRISTOL MYERS SQUIBB CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Separating a desired protein from a mixture of components supplied to cells, cellular by-products, and aggregated forms of the protein to sufficient purity (e.g., sufficient to be useful as a human therapeutic) presents formidable challenges to biologics manufacturers

Method used

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  • Methods of measuring hydrophobicity of chromatographic resin
  • Methods of measuring hydrophobicity of chromatographic resin
  • Methods of measuring hydrophobicity of chromatographic resin

Examples

Experimental program
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preparation example Construction

[0069] Preparation of the mixture depends on the initial protein expression. Some cell systems direct the protein (e.g., antibody) secreted from the cell into the surrounding growth medium, other systems will be retained within the cell antibodies. For proteins produced in the cell, you can use a variety of methods (such as mechanical shear, osmotic shock and enzymatic treatment) in any one of disrupted cells. Destruction of the cells so that all the contents released into the homogenate, and in addition produce subcellular debris, the debris can be removed by centrifugation or filtration. In the process of protein production run, since the natural death of cells released host cell proteins and intracellular proteins directly secreted similar problems occur, although to a lesser extent.

[0070] In one embodiment, the cells or cell debris is removed from the mixture, for example, to prepare a clear body. The method of the present invention may use any suitable method to remove cel...

Embodiment 1

[0074] The method described herein is a hydrophobic integrally with a fluorescent dye technique gold nanoparticles functionalized characterized chromatography resins. Currently, small globular model protein (e.g., ribonuclease A, lysozyme) of retention time measurement is relatively hydrophobic adsorbent standard methods, especially for hydrophobic interaction chromatography (HIC) resin (GE Healthcare, Certificate of Analysis of Phenyl Sepharose TM 6Fast Flow (high sub), test and limit: AS 45-6003-91Ed.AG 3). However, for a wide range of types of chromatography, general method for quantifying the hydrophobic resin it is still lacking and is very desirable. Has been used for measuring surface hydrophobic fluorescent dye characterized cation exchange chromatography (the CEX) resin aggregate (lumped) hydrophobic properties (Chen Z, Huang C, Chennamsetty N, Xu X, Li ZJ.JChromatogr A.2016,1460 : 110-122). Due to the small size of dye used (typically 10,000 daltons). In addition, the c...

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Abstract

In certain embodiments, the present invention provides a method of measuring the level of hydrophobicity of a chromatographic resin. In certain embodiments, the present invention provides a method of selecting a chromatographic resin condition for purifying a protein of interest from a mixture, wherein the protein of interest has low or no aggregation formation during chromatography. In certain embodiments, the present invention provides a method of selecting the chromatographic resin from a plurality of chromatographic resins for purifying the protein of interest from a mixture, wherein the protein of interest has low or no aggregation formation during chromatography.

Description

[0001] Cross-reference related application [0002] This application claims the benefit of US Provisional Application No. 29 March 2019 filed equity 62 / 826,260, and which is incorporated herein by reference in its entirety. Background technique [0003] Economic purification of proteins is large-scale biopharmaceutical industry increasingly important issue. Therapeutic proteins are typically prokaryotic or eukaryotic cell line, the cell line was engineered to expressed from a recombinant plasmid containing the gene encoding a protein of the protein. Aggregates byproduct cells, and proteins isolated from a mixture of components supplied to the cells to a sufficient desired purity of the protein (e.g., sufficient for use as human therapeutics) manufacturers of biological formulations presents formidable challenge. [0004] Thus, in the art a need for improved methods of protein purification that can be used to accelerate the large scale processing of Protein-based therapeutics (suc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/89B01D15/38B01J20/286C07K1/18
CPCG01N30/89B01D15/3804C07K1/18C07K1/20C07K1/16B01D15/00B01D15/327B01D15/361B01D15/3847B82Y30/00G01N31/22G01N33/442B01D15/362B82Y35/00B82Y40/00G01N30/50G01N2030/027
Inventor 黄弨J·J·佩里徐晅阔S·高斯李正剑W·金
Owner BRISTOL MYERS SQUIBB CO