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 convectionComparison 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™ Monolithic Resins and Columns

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™ technologyDuloCore™ technology

DuloCore™ Monolithic Resins

Product Name Pore Size Categories
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

DuloCore™ Monolithic Columns

Product Name Pore Size Categories
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™ Resin and Column Specifications

DuloCore™ Matrix QA DEAE SO3
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
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)

Characteristics and Performance Features

High Operational Flow Rates in Convection Mode
  • Operating linear velocity of up to 3,000 cm/hr and backpressure of less than 0.2 Mpa, which enables superior throughput capacity relative to competitors
Adjustable Resin Pore Size and Particle Size
  • High recovery rate and purity for viral vectors or large biomolecules
High Reproducibility
  • Lot-to-lot consistency of viral capture performance
Easy Scalability
  • Polymer based resins
  • Easy to pack in any sizes of commercial columns
Durability
  • Excellent chemical stability for multiple CIPs cycles
  • Good pH stability
  • Excellent bed stability of industrial scale columns under high flow rates
Cost-effectiveness
  • Economic pricing for single use
Quality
  • DuloCore™ resin factory certified by ISO 9001:2015 Quality Management System and cGMP

Applications

Rapid and High Yield Purification of Plasmid DNA by DuloCore™ DEAE

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 purificationHigh purity of plasmid DNA from DuloCore™ DEAE purification

Separation of Empty and Full AAV8 Capsids Using DuloCore™ QA

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 methodThe chromatogram and TEM for empty and full AAV8 capsids separation by step elution method

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