A More Efficient Chromatography Mode-MMC

Published by News Editor on

MMC Overview

Mixed-mode chromatography(MMC) optimized the structure of the functional ligand, combining two or more interaction modes to enhance affinity and selectivity to the target protein, thus improving purification efficiency. Compared with the traditional IEC and HIC, the new mode MMC has advantages of high loading capacity, high selectivity and high efficiency.

MMC Features

  1. It has many types of interactions with target molecules, including ionic interaction, hydrogen bonding and hydrophobic interaction.
  2. The MMC medium has a high ligand density and non-salt-dependent adsorption properties. Efficient protein elution can be realized by adjusting pH.
  3. Protein can interact with hydrophobic groups and ion exchange groups coupled by the MMC media: electrostatic interaction plays a dominant role when ion strength is weak; Hydrophostatic interaction plays a dominant role when ion strength is strong; Hydrophobicity and static electricity may interact together when ion strength is medium.
  4. Protein elution usually has two ways: when hydrophobic binding plays a dominant role, it can be realized by reducing the pH or taking advantage of the electrostatic rejection between proteins and ligand. When electrostatic binding plays a dominant role, it can be realized by enhancing the ion strength and reducing the electrostatic interaction.

There are two composite ligands, MMC and MIX A, technical parameters are shown as below:

Solid MMC

NameSolid MMC
Bead structureHighly rigid agarose
Bead size range45-120μm
Mean particle size75μm
TypeWeak cation exchange
Ionic capacity70-90 μmol H+/mL
pH stability3-12 (long term), 2-14 (short term)
Operating pressure≤0.5MPa
Flow rate90-500cm/h
Chemical stabilityAll common buffers, 1M NaOH, 8M urea, 6M guanidine hydrochloride,70% ethanol, avoid using oxidants and anionic detergent
Storage solvent20% ethanol
Storage temperature4-30℃

Application

Column: TXK 50/30

Medium type: Solid MMC

Sample: 300M Fermentation broth of a recombinant pichia pastoris Equilibrium buffer: 0.4 M NaCl, 20 mM PB, pH 6.5

Eluting buffer: 1.5 M NaCl,20 mM PB, pH 6.5

Eluting method: gradient elution

Solid MIX A

NameSolid MIX A
Bead structureHighly rigid agarose
Bead size range45-120μm
Mean particle size75μm
Type Strong anion exchange
Ionic capacity90-120 μmol Cl/ml
pH stability        4-12 (long term), 2-14 (short term)
Operating pressure≤0.5MPa
Flow rate90-500cm/h
Chemical stabilityAll common buffers, 1M NaOH, 1M HAc, 8M urea, 6M guanidine hydrochloride, 70% ethanol, avoid using oxidants, anionic detergent
Storage solvent20% ethanol
Storage temperature4-30℃

Application

Column: TXK 50/30

Medium type: Solid MiX A

Sample: 300M Fermentation broth of a recombinant pichia pastoris

Equilibrium buffer: 0.1 M NaCl, 20 mM phosphate buffer, pH 7.0

Eluting buffer: 1.5 M NaCl, 20 mM phosphate buffer, pH 7.0

 Solid MMC Mustang & Solid MIX A Mustang

Name       Solid MMC Mustang                       Solid MIX A Mustang
Bead structure           Highly rigid agarose
Bead size range25-45μm
Mean particle size34μm
TypeComposite weak cation exchange Composite strong anion exchange
Ionic capacity   25-39 μmol H+/ml               80-110 μmol Cl-/ml
pH stability4-12(long term), 2-14(short term)
Operating pressure≤0.5MPa
Flow rate90-180cm/h
Chemical stabilityAll common buffers, 1M NaOH, 1M HAc, 8M urea, 6M guanidine hydrochloride, 70% ethanol, avoid using oxidants, anionic detergent.
Storage solvent20% ethanol
Storage temperature4-30℃

Solid MIX A Mustang

P/NSpecification
IMNH0125ml
IMNH02100ml
IMNH03500ml
IMNH041L

Solid MMC Mustang

P/NSpecification
IMPH0125ml
IMPH02100ml
IMPH03500ml
IMPH041L

If you have any problem or require further information, please contact info@welchmat.com.

Categories: News

0 Comments

Leave a Reply

Your email address will not be published. Required fields are marked *