Method for enhancing dynamic absorption volume of ion exchange absorbent of transverse electric field

A technology of ion exchange adsorption and dynamic adsorption, applied in ion exchange, ion exchange regeneration, ion exchange treatment devices, etc., can solve the problems of multiple chambers, complex structure, increased distance between electrodes, etc., and achieve low price and wide application Prospect, effect of improving adsorption capacity

Inactive Publication Date: 2007-06-13
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the structure of the device is more complicated and there are many chambers
The cavity of the middle filling material is very thin, and if the cavity is enlarged, the distance between the two electrodes will increase

Method used

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  • Method for enhancing dynamic absorption volume of ion exchange absorbent of transverse electric field

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Experimental program
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Effect test

Embodiment 1

[0019] Dynamic adsorption curves of bovine serum albumin on DEAE-Sepharose FF under different electric field intensities. Sodium 3.9mmol / L tris-47mmol / L glycine buffer (pH8.2). The eluent and regeneration solution were 3.9mmol / L tris-47mmol / L glycine buffer (pH8.2) containing 0.5 and 1.0mol / L sodium chloride, respectively. The concentration of the BSA sample solution was 2.0 mg / mL. During the experiment, the electrode solution was cooled at 6 °C by a cooler, and the mobile phase was cooled by an ice-water mixture.

[0020] Load the pre-equilibrated DEAE-Sepharose FF into the packing chamber by gravity settling. Prior to sample loading, the packing chamber was equilibrated with equilibration buffer until the UV (280nm) absorbance stabilized around the baseline. Then, apply an alternating electric field on both sides of the gel column, and add the pre-cooled BSA sample solution at a constant flow rate. The concentration of protein at the outlet is detected by online UV, and ...

Embodiment 2

[0022] Use DEAE-Sepharose FF to separate bovine serum albumin and immunoglobulin G mixture. The size of the electrochromatographic packing chamber is (length×width×depth) 3.0×0.5×1.2cm, and the equilibrium buffer contains 5.0mmol / L sodium chloride 3.9mmol / L tris-47mmol / L glycine buffer solution (pH8.2); eluent and regeneration solution are 3.9mmol / L trimethylol containing 0.5 and 1.0mol / L sodium chloride respectively Aminomethane-47mmol / L glycine buffer (pH8.2). The protein mixture with a total concentration of 2.0 mg / mL is prepared by mixing equal volumes of bovine serum albumin and immunoglobulin G sample solutions with a concentration of 2.0 mg / L. During the experiment, the electrode solution was cooled at 6 °C by a cooler, and the mobile phase was cooled by an ice-water mixture.

[0023] Pre-equilibrated packing (DEAE-Sepharose FF) was loaded into the central chamber by gravity settling. The electrode voltage was 75 volts, and the current change period was 20 seconds. A...

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Abstract

The invention discloses a method for improving the fluidizing adsorption capacity of the biomacromolecule such as protein on the ion exchange sorbent with an alternating electric field. Said preparative electrochromatogram separation device comprises a gel chamber and electrode chambers at double- side. The method for separation of biomacromolecule with this device is characterized in that: acetate, phosphonate, hydrogen carbonate , or trishydroxymethylaminomethane- hydrochloric acid buffer liquid is prepared as a mobile phase, the flow velocity of the mobile phase being 10- 200cm / h and the temperature being 2- 25Deg. C; iso- period alternating electric field with a voltage of 1- 600V and a current alternating period of 2- 200 seconds is stressed at the two sides of gel chamber, the flow velocity of electrode liquid being 20- 400cm / h and the temperature being 4- 25 Deg. C; and the specimen solution is sampled by means of meeting head and the sampling is stopped when the penetration approaches among 2- 80% of the sample concentration. Invented is characterized in that the separating power is strong, the handling capacity is large, the applicability extensive, and it has an extensive application prospect in separating and purifying the biomacromolecule such as protein and nucleic acid.

Description

technical field [0001] The invention relates to a method for improving the dynamic adsorption capacity of biomacromolecules such as protein on an ion exchange adsorbent by using an alternating electric field, and belongs to the technical field of biological product processing and separation. Background technique [0002] At present, in the processing of biological downstream products, chromatographic separation has an irreplaceable position by other separation methods. However, the pore size of most chromatographic media is on the order of nanometers, and the mass transfer coefficient of proteins in the above media is 2-3 orders of magnitude lower than that of small molecules, which seriously affects the mass transfer of biological macromolecules in the chromatographic media. Therefore, strengthening the internal mass transfer of the medium and improving its separation efficiency for processing biomacromolecules is one of the urgent problems to be solved in chromatography se...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/08C07K1/18
Inventor 孙彦谭国民史清洪董晓燕白姝
Owner TIANJIN UNIV
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