Porous particles with different size, porosity and chemical composition are used as chromatography column packing for the separation, isolation and/or purification of biological compounds. Resins are classic chromatography media. Traditional resin beads have a porous structure that contain micro-pores where the binding sites are located. Solutes are carried to sites on the particle surface by bulk convective flow of the mobile phase through the resin column and then diffuse to binding surfaces inside the particle. The intraparticle diffusion process may be quite slow for the large molecules. To maximize binding capacity and resolution, interaction with as many sites as possible is necessary, which is mostly relying on the diffusion effect of large molecules in the resin bead media.
Monolithic columns consist of one piece of a continuous, porous material that fills the column entirely. Within this monolith, the continuous skeleton is filled with inter-connected pores that form flow channels of uniform size. The interphase mass transfer in monoliths is governed by fluid convection. The monolithic column develops a network of channels in the continuous phase of the porous material, showing higher axial permeability, larger internal pore surface area, and less back pressure than that of traditional packed columns. These channels allow better contact between the analyte and the active site of the stationary phase. Therefore, these monoliths can be used for biomolecular separation, especially for large biological particles and allow performing separation process at high flow rates and low back pressures.
Comparison of chromatographic columns using fluid diffusion and fluid convection
Amerigo Scientific offers DuloCore™ monolithic resins, a combination of the benefits of bead packing materials and monolithic columns. DuloCore™ monolithic resins can provide high binding capacity for large biological particles and use fluid convection mode to achieve excellent separation performance at high flow rate and low back-pressure, with easy scale-up, non-clogging, economical price, high recovery, mechanical strength and chemical resistance.
DuloCore™ spherical matrix is a cross-linked polymethacrylate material specifically designed to purify large biomolecules such as plasmid DNAs, viruses, mRNAs, exosomes, proteins, peptides, and more. Traditional resin beads are more suitable for protein and antibody purification (about 10 nm) due to their small pore structure. Unlike traditional resin beads, which use fluid diffusion in the pores to separate molecules, DuloCore™ resins have large inter-connected pores that form flow channels in resins, showing excellent mechanical strength in high flow convection mode. Therefore, DuloCore™ monolithic resins are particularly suitable for purification of biological macromolecular particles in mass production. DuloCore™ resins with pore sizes of 1.0 μm or 1.6 μm can maintain low back pressure at ultra-high flow rates of 2400 cm/hr without bed collapse problems, which is attributed to the excellent mechanical strength and monolithic macroporous structure of DuloCore™.
DuloCore™ technology
Product Name | Pore Size | Categories |
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TANTTI®DuloCore™ Resin (1.0μm) | 1.0 μm | - |
TANTTI®DuloCore™ Resin (1.6μm) | 1.6 μm | - |
TANTTI®DuloCore™ Q (1.0μm) | 1.0 μm | Ion exchange chromatography, strong anion exchange (SAX) |
TANTTI®DuloCore™ Q (1.6μm) | 1.6 μm | Ion exchange chromatography, strong anion exchange (SAX) |
TANTTI®DuloCore™ S (1.0μm) | 1.0 μm | Ion exchange chromatography, strong cation exchange (SCX) |
TANTTI®DuloCore™ S (1.6μm) | 1.6 μm | Ion exchange chromatography, strong cation exchange (SCX) |
TANTTI®DuloCore™ DEAE (1.0μm) | 1.0 μm | Ion exchange chromatography |
TANTTI®DuloCore™ DEAE (1.6μm) | 1.6 μm | Ion exchange chromatography |
TANTTI®DuloCore™ C4 (1.0μm) | 1.0 μm | Hydrophobic interaction chromatography |
TANTTI®DuloCore™ C4 (1.6μm) | 1.6 μm | Hydrophobic interaction chromatography |
TANTTI®DuloCore™ Oligo dT (1.0μm) | 1.0 μm | Affinity chromatography |
TANTTI®DuloCore™ Oligo dT (1.6μm) | 1.6 μm | Affinity chromatography |
Product Name | Pore Size | Categories |
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TANTTI®DuloCore™ Q (1.0μm), Pre-Packed Column | 1.0 μm | Ion exchange chromatography, strong anion exchange (SAX) |
TANTTI®DuloCore™ Q (1.6μm), Pre-Packed Column | 1.6 μm | Ion exchange chromatography, strong anion exchange (SAX) |
TANTTI®DuloCore™ S (1.0μm), Pre-Packed Column | 1.0 μm | Ion exchange chromatography, strong cation exchange (SCX) |
TANTTI®DuloCore™ S (1.6μm), Pre-Packed Column | 1.6 μm | Ion exchange chromatography, strong cation exchange (SCX) |
TANTTI®DuloCore™ DEAE (1.0μm), Pre-Packed Column | 1.0 μm | Ion exchange chromatography |
TANTTI®DuloCore™ DEAE (1.6μm), Pre-Packed Column | 1.6 μm | Ion exchange chromatography |
TANTTI®DuloCore™ C4 (1.0μm), Pre-Packed Column | 1.0 μm | Hydrophobic interaction chromatography |
TANTTI®DuloCore™ C4 (1.6μm), Pre-Packed Column | 1.6 μm | Hydrophobic interaction chromatography |
TANTTI®DuloCore™ Oligo dT (1.0μm), Pre-Packed Column | 1.0 μm | Affinity chromatography |
TANTTI®DuloCore™ Oligo dT (1.6μm), Pre-Packed Column | 1.6 μm | Affinity chromatography |
DuloCore™ | Matrix | QA | DEAE | SO3 |
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Functional Group | Polymethacrylic based resin | Quaternary amine | Diethylaminoethyl | Sulfonate |
Epoxide | strong anion exchanger | weak anion exchanger | strong cation exchanger | |
Dynamic Binding Capacity (BSA)* | ND | ≧12 mg/mL (1.6 μm) ≧40 mg/mL (1.0 μm) |
≧14 mg/mL (1.6 μm) | ≧25 mg Lysozyme /mL |
Dynamic Binding Capacity* (Thyroglobulin) | ND | ≧10.2 mg/mL (1.6 μm) ≧9.5 mg/mL (1.0 μm) |
≧25 mg/mL (1.6 μm) | ND |
Polymer Matrix | Polymethacrylate | |||
Average particles size (D50) | 50 ± 10 μm | |||
Particles size distribution (D90-D10) | 34.3 ± 5.9 μm | |||
Average inter-pore size | 1.0 ± 0.15 μm; 1.6 ± 0.15 μm | |||
Epoxy Content (μmol/mL) | 0.9 - 1.2 | ND | ||
Ionic capacity (μmol/mL) | ND | 50 – 100 | 20-70 | |
Column Volume | 50 ml, 100 ml, 500 ml, 1 L, 5 L |
1 mL (7 mm ID X 2.5 cm) 5mL (16 mm ID X 2.5 cm) 50 ml, 100 ml, 500 ml, 1 L, 5 L |
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Housing | HDPE | Polypropylene | ||
Housing Max. pressure | 0.5 MPa | |||
Operating Flow Rates | 0.1 - 20 mL/min | |||
Connector | 10-32 UNF coned port for 1/16” OD tubing connection | |||
Recommended pH | pH 2 – 13 | pH 2 – 10 | pH 2 – 13 | |
Storage temperature | 4 °C | 4 °C - 30 °C | ||
Storage | 2 years | |||
Shelf Life | 20% ethanol |
*DBC: BSA and Thyroglobulin (Flow rate: 300 cm/h)
High Operational Flow Rates in Convection Mode |
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Adjustable Resin Pore Size and Particle Size |
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High Reproducibility |
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Easy Scalability |
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Durability |
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Cost-effectiveness |
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Quality |
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As shown in the figure below, DuloCore™ DEAE column demonstrated its separation capabilities. In linear salt gradient elution, the elution peaks of plasmid DNA (7.3 kb) and protein impurities were easily observed. The purity of the DNA fractions was confirmed using gel electrophoresis. Subsequently, the flow rate (1 to 4 mL/min) and sample loading (1 to 20 mL) in DNA purification process were optimized.
High purity of plasmid DNA from DuloCore™ DEAE purification
As shown in the figure below, DuloCore™ QA column demonstrated its capability to separate empty AAV8 capsids from the full AAV8 capsids through simple isocratic elution. The full AAV8 capsids could be recovered at higher conductivity.
The chromatogram and TEM for empty and full AAV8 capsids separation by step elution method
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