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
- It has many types of interactions with target molecules, including ionic interaction, hydrogen bonding and hydrophobic interaction.
- The MMC medium has a high ligand density and non-salt-dependent adsorption properties. Efficient protein elution can be realized by adjusting pH.
- 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.
- 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
| Name | Solid MMC |
| Bead structure | Highly rigid agarose |
| Bead size range | 45-120μm |
| Mean particle size | 75μm |
| Type | Weak cation exchange |
| Ionic capacity | 70-90 μmol H+/mL |
| pH stability | 3-12 (long term), 2-14 (short term) |
| Operating pressure | ≤0.5MPa |
| Flow rate | 90-500cm/h |
| Chemical stability | All common buffers, 1M NaOH, 8M urea, 6M guanidine hydrochloride,70% ethanol, avoid using oxidants and anionic detergent |
| Storage solvent | 20% ethanol |
| Storage temperature | 4-30℃ |
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
| Name | Solid MIX A |
| Bead structure | Highly rigid agarose |
| Bead size range | 45-120μm |
| Mean particle size | 75μm |
| Type | Strong anion exchange |
| Ionic capacity | 90-120 μmol Cl–/ml |
| pH stability | 4-12 (long term), 2-14 (short term) |
| Operating pressure | ≤0.5MPa |
| Flow rate | 90-500cm/h |
| Chemical stability | All common buffers, 1M NaOH, 1M HAc, 8M urea, 6M guanidine hydrochloride, 70% ethanol, avoid using oxidants, anionic detergent |
| Storage solvent | 20% ethanol |
| Storage temperature | 4-30℃ |
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 range | 25-45μm |
| Mean particle size | 34μm |
| Type | Composite weak cation exchange Composite strong anion exchange |
| Ionic capacity | 25-39 μmol H+/ml 80-110 μmol Cl-/ml |
| pH stability | 4-12(long term), 2-14(short term) |
| Operating pressure | ≤0.5MPa |
| Flow rate | 90-180cm/h |
| Chemical stability | All common buffers, 1M NaOH, 1M HAc, 8M urea, 6M guanidine hydrochloride, 70% ethanol, avoid using oxidants, anionic detergent. |
| Storage solvent | 20% ethanol |
| Storage temperature | 4-30℃ |
Solid MIX A Mustang
| P/N | Specification |
| IMNH01 | 25ml |
| IMNH02 | 100ml |
| IMNH03 | 500ml |
| IMNH04 | 1L |
Solid MMC Mustang
| P/N | Specification |
| IMPH01 | 25ml |
| IMPH02 | 100ml |
| IMPH03 | 500ml |
| IMPH04 | 1L |
If you have any problem or require further information, please contact info@welchmat.com.
0 Comments