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WorkBeads™ 40/1000 ACT and WorkBeads 40/10 000 ACT pre-activated separation resins are based on highly cross-linked agarose. The resins can be used for stable covalent coupling of proteins, peptides or low-molecular weight substances exposing primary amine or thiol groups. The purpose of the ligand coupling may be to create an affinity resin or an immobilized-enzyme reactor.
- Ideal for coupling of specific customer designed resins
- Stable covalent linkage
- Suitable for coupling of ligands containing thiol, amino and hydroxyl groups
- Two different porosities for optimized results
Product
Agarose based beads have been successfully used for decades in life sciences research and for industrial purification of proteins, peptides, oligonucleotides and other biomolecules. This compatibility depends on the hydrophilic nature and minimal unspecific interactions between agarose and these compounds.
WorkBeads 40/1000 ACT and WorkBeads 40/10 000 ACT contain reactive bromohydrin groups. Coupling can be done under neutral or slightly basic conditions in aqueous solution. These activated resins are stable in the absence of amines and thiols, and can be stored in 20% ethanol at room temperature until use. The nature of this chemistry makes coupling easy and convenient. WorkBeads 40/1000 ACT and WorkBeads 40/10 000 ACT have different porosities, which will affect the amount of ligand that can be coupled and the binding capacity of a target substance binding to the ligand, i.e., the prepared affinity resin. As rule of thumb; WorkBeads 40/1000 ACT should be used for small protein ligands or small affinity targets, and WorkBeads 40/10 000 ACT should be used for large protein ligands and/or large affinity targets. An affinity resin prepared by coupling a stable ligand on these active resin can be used and/or stored for several years.
Table 1. Main characteristics of WorkBeads 40/1000 ACT and WorkBeads 40/10 000 ACT resins
|
WorkBeads 40/1000 ACT |
WorkBeads 40/10 000 ACT |
|
|
Target substances |
Small molecules and peptides |
Small molecules and peptides, proteins, e.g., Immunoglobulins |
|
Target groups |
Thiol, amino, and hydroxyl groups |
Thiol, amino, and hydroxyl groups |
| Matrix |
Rigid, highly cross-linked agarose |
Rigid, highly cross-linked agarose |
| Average particle size¹ (Dv50) | 45 µm | 45 µm |
|
Reactive groups |
Bromohydrin |
Bromohydrin |
|
Exclusion limit |
1 x 106 Da (globular proteins) |
1 x 107 Da (globular proteins) |
|
Max flow rate² |
600 cm/h | 600 cm/h |
|
Reactive groups content |
200 µmol/ml |
200 µmol/ml |
|
Chemical stability (before coupling³) |
Buffers pH < 8.5 |
Buffers pH < 8.5 |
|
Chemical stability (after coupling⁴) |
Compatible with all standard aqueous buffers used for protein purification, 1 M NaOH, 30% isopropanol or 70% ethanol. Should not be stored at < pH 3 for prolonged time |
Compatible with all standard aqueous buffers used for protein purification, 1 M NaOH, 30% isopropanol or 70% ethanol. Should not be stored at < pH 3 for prolonged time |
|
pH stability⁴ |
2 – 13 (after coupling) | 2 – 13 (after coupling) |
| Storage⁵ | 2 to 25 °C in 20% ethanol | 2 to 25 °C in 20% ethanol |
¹ The median particle size of the cumulative volume distribution.
² Determined in water using a 10 x 300 mm column.
³ Avoid substances containing thiol and amino groups. Substances containing hydroxyl groups Will only react if deprotonated. The unreacted resin is generally stable in alcohols at neutral PH.
⁴ Agarose matrix and linker. Stability of the coupled substance may differ.
⁵ The choice of storage conditions for the coupled resin depends on the nature of the ligand.