Optimization of elution salt concentration in stepwise elution of protein chromatography using linear gradient elution data

2006 ◽  
Vol 1114 (1) ◽  
pp. 97-101 ◽  
Author(s):  
Takashi Ishihara ◽  
Toshihiko Kadoya ◽  
Naomi Endo ◽  
Shuichi Yamamoto
Keyword(s):  
Salt Concentration ◽  
Gradient Elution ◽  
Linear Gradient ◽  
Protein Chromatography

scholarly journals A simple one-step procedure for the separation of calpain I, calpain II and calpastatin

1985 ◽  
Vol 231 (1) ◽  
pp. 201-204 ◽  
Author(s):  
J O Karlsson ◽  
S Gustavsson ◽  
C Hall ◽  
E Nilsson
Keyword(s):  
Salt Concentration ◽  
Soluble Fraction ◽  
Rabbit Brain ◽  
Endogenous Inhibitor ◽  
Linear Gradient ◽  
Step Procedure ◽  
Neuronal Proteins ◽  
One Step ◽  
Membrane Bound ◽  
Calpain Ii

The soluble fraction from rabbit brain was adsorbed on a column of phenyl-Sepharose. By applying a linear gradient with decreasing salt concentration and increasing pH, it was possible to separate calpain I and calpain II from each other and from the endogenous inhibitor calpastatin. Both enzymes were capable of degrading endogenously labelled neuronal proteins, including slowly axonally transported soluble proteins and rapidly transported membrane-bound proteins, as well as casein.

scholarly journals A method for designing flow-through chromatography processes

2019 ◽  
Vol 268 ◽  
pp. 01004
Author(s):  
Noriko Yoshimoto ◽  
Sumiko Hasegawa ◽  
Shuichi Yamamoto
Keyword(s):  
Membrane Filtration ◽  
Continuous Process ◽  
Gradient Elution ◽  
Downstream Processing ◽  
Stable Operation ◽  
Continuous Manufacturing ◽  
Linear Gradient ◽  
Unit Operations ◽  
Flow Through ◽  
Small Change

Continuous manufacturing is expected to increase the productivity of the production of biologics such as protein drugs. However, it is not easy to build the continuous process especially for downstream processing as many unit operations (chromatography and membrane filtration) are involved. An operation method known as flow-through chromatography (FTC) is considered to be an efficient method for separating two components as the flow is continuous. In FTC, a target protein is eluted from the chromatography column without adsorption whereas contaminants are strongly bound. Since at least two different modes of chromatography are needed in order to remove contaminants, two FTC columns have to be connected in order to build the continuous process. This is not an easy task since the mobile phase properties (pH, salt, buffer ions) are different for the two columns. In this paper, we developed a method for designing FTC processes based on the mechanistic models that we have developed for linear gradient elution (LGE) of proteins. The distribution coefficient K as a function of salt concentration, I determined from LGE data were used for the simulation. It was found that the process is quite sensitive to a small change in I. This indicates that FTC processes are carefully monitored and controlled for the stable operation.

A new flow-injection chromatography method exploiting linear-gradient elution for fast quantitative screening of parabens in cosmetics

2016 ◽  
Vol 8 (47) ◽  
pp. 8337-8344 ◽  
Author(s):  
Margarita Barbatsi ◽  
Michael Koupparis ◽  
Anastasios Economou
Keyword(s):  
Flow Rate ◽  
Flow Injection ◽  
Gradient Elution ◽  
Chromatography Method ◽  
Linear Gradient ◽  
Rate Modulation ◽  
Quantitative Screening

Linear gradient elution flow injection chromatography with flow rate modulation was used for the determination of 4 parabens.

scholarly journals Bioassay-Guided Separation of Centipeda minima Using Comprehensive Linear Gradient Centrifugal Partition Chromatography

Molecules ◽  
2020 ◽  
Vol 25 (13) ◽  
pp. 3077
Author(s):  
Ji Hoon Kim ◽  
Eun Ju Jung ◽  
Yun Jung Lee ◽  
En Mei Gao ◽  
Ahmed Shah Syed ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Gradient Elution ◽  
Solvent System ◽  
Luciferase Assay ◽  
Partition Chromatography ◽  
Centrifugal Partition Chromatography ◽  
Lower Phase ◽  
Linear Gradient ◽  
Centipeda Minima ◽  
Water Saturated

A comprehensive linear gradient solvent system for centrifugal partition chromatography (CPC) was developed for the bioassay-guided isolation of natural compounds. The gradient solvent system consisted of three different ternary biphasic solvents types: n-hexane–acetonitrile–water (10:2:8, v/v), ethyl acetate–acetonitrile–water (10:2:8, v/v), and water-saturated n-butanol–acetonitrile–water (10:2:8, v/v). The lower phase of the n-hexane–acetonitrile–water (10:2:8, v/v) was used as the stationary phase, while its upper phase, as well as ethyl acetate–acetonitrile–water (10:2:8), and water-saturated n-butanol–acetonitrile–water (10:2:8, v/v) were pumped to generate a linear gradient elution, increasing the mobile phase polarity. We used the gradient CPC to identify antioxidant response elements (AREs), inducing compounds from Centipeda minima, using an ARE-luciferase assay in HepG2 cells, which led to the purification of the active molecules 3-methoxyquercetin and brevilin A. The developed CPC solvent systems allow the separation and isolation of compounds with a wide polarity range, allowing active molecule identification in the complex crude extract of natural products.

Influence of the pre-elution of solute in initial mobile phase on retention time and peak compression under linear gradient elution

2020 ◽  
Vol 1618 ◽  
pp. 460858 ◽  
Author(s):  
Weiqiang Hao ◽  
Kai Wang ◽  
Bangyi Yue ◽  
Qiang Chen ◽  
Yibo Huang ◽  
...  
Keyword(s):  
Retention Time ◽  
Mobile Phase ◽  
Gradient Elution ◽  
Linear Gradient ◽  
Peak Compression
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