Protein Technologies – Bioseparation Products
Mimetic Ligand™ Adsorbents: Other Mimetics™
MIMETIC ligands™ are highly stable, synthetic affinity ligands which are capable of interacting with a wide range of proteins. Unlike the crude textile dyes which have been used previously, MIMETIC ligands™ have purpose-designed structures which are synthesized and immobilized to solid supports by extremely stable spacer-arm linkages. They offer higher degrees of purification, minimal ligand leakage and more reproducible separations than can be achieved by low quality textile dye adsorbents. A diverse range of ligand structures have been developed, each of which incorporates features which mimic the protein-binding components of natural ligands. Consequently each MIMETIC ligand™ has a very different specificity allowing a wide range of proteins to be purified.
Mimetic™ Adsorbents: Properties
- ROBUSTNESS - Mimetic Ligands™ are biologically inert, very durable, have excellent liquid flow properties, and are resistant to most chemical treatments. Mimetic Ligand™ adsorbents are stable in the pH range 2 - 14 and can be treated with denaturants (8M urea), detergents (Triton, SDS) and can be sterilized by autoclaving.
- DEFINED STRUCTURE - Each ligand structure has been designed and synthesized in-house and is of known purity. These factors allow precise control of immobilized ligand concentration providing accurate batch reproducibility and hence chromatographic reproducibility.
- HIGH CAPACITIES - In general, Mimetic Ligand™ adsorbent capacities are higher than other affinity adsorbents. Depending upon applied sample purity, type of protein and operational condition, capacities can exceed 50mg protein per ml of packed gel.
- LOW LIGAND LEAKAGE - Mimetic Ligands™ are immobilized by a highly stable spacer-arm linkage which eliminates detectable ligand leakage across a wide pH range.
- HIGH RECOVERIES - Protein absorption and desorption may be performed at physiological pH without the need for denaturants leading to high recoveries of activity.
Mimetic Ligand™ Adsorbents: Advantages
- UNIVERSALLY APPLICABLE - The Mimetic Ligand™ adsorbent range has been designed to enable purification of many proteins, providing a general technique that can replace ion-exchange, gel permeation and hydrophobic interaction chromatography, but with higher recovery and purity.
- UNKNOWN OR UN-CHARACTERIZED TARGET PROTEINS - Mimetic Ligand™ adsorbents can be used without detailed information on the target protein (e.g. molecular weight or iso-electric point).
- BI-MODAL PURIFICATION - Purification may be achieved by positive binding (target protein bound; contaminants un-retarded) or negative binding (contaminants bound; target protein un-retarded).
- ALKALI-STABLE ADSORBENTS - Enables cleaning, sterilization and depyrogenation with 1M sodium hydroxide, ensuring long operational lifetimes.
(Note: Reversible color changes at high pH may be observed in some instances but the chromatographic properties and protein binding capacity of the adsorbents are unaffected. Mimetic Ligand adsorbents are stable indefinitely in the pH range 3 - 13 and may be treated intermittently with solutions in the range pH 2 - 14. Ligand leakage is less than 5 X 1010 Mol/ml throughout this pH range.)
Specifications
| Support Matrix: | 6% cross-linked agarose |
| Bead Size: | 45 - 165 μm |
| Exclusion Limit (Globular Proteins): | 4 x 106 |
| Immobilized Ligand Concentration: | 2.5 +/-0.3 μmol.g-1 moist weight |
| (36 +/-4 μmol.g-1 dry weight) | |
| Temperature Range: | 4 - 120° C |
| рH Range: | 3 - 13 (continuous) |
| 2 - 14 (intermittent) | |
| Solvent Compatibility: | Compatible with alcohols, esters, ketones and ethers |
| Protein Binding Capacity: | Can exceed 40mg protein per ml packed gel |
| Recommended Preservative: | Ethanol /0.1 M NaCl [25:75 v/v] |
| Cleaning/Sanitization/Depyrogenation: | 1M NaOH |
| Sterilization: | 1M NaOH, or autoclave 30min. at 120°C, pH 7.0 |
| Storage: | 4°C in preservative |
Mimetic Ligand™ affinity adsorbents are compatible with most biological buffers and commonly encountered buffer additives, including:
- Chaotropes (e.g. 8M urea; 6M guanidine HCl)
- Detergents (inc. SDS and Triton X-100)
- Chelating agents (e.g. EDTA)
- Thiols (e.g. 2-mercaptoethanol; DTT)
Mimetic Ligand™ adsorbents are compatible with metal ions but some discoloration (persisting after treatment with 1M NaOH) may be observed following contact with certain di- and tri- valent metal ions (e.g. Cu2+, Co2+). Multivalent metal ions can influence the chromatographic properties of Mimetic Ligand™ adsorbents and they may be added deliberately to enhance protein binding. Bound metal ions are readily desorbed by treatment with 5mM EDTA.
Mimetic Ligand™ adsorbents may be sterilized by autoclaving for 30mins. at 120°C in 50mM sodium phosphate buffer, pH 7.0. Autoclaving may cause very slight ligand leakage due to low level dissolution of the agarose matrix. However, the protein binding capacity of the adsorbent is unaffected by this treatment.
Flow Rate Performance of Mimetic Blue 1 A6XL
Using Mimetic Ligand™ Adsorbents
Mimetic Ligand™ affinity adsorbents may be used at any stage in a purification process and may be used repeatedly even when regenerated with harsh chemical treatments. Purification may be achieved in a positive mode whereby the target protein is bound and contaminants pass through the adsorbent bed, or else in a negative mode, where contaminating proteins are bound and the target protein is unretarded. A combination of one negative binding step followed by one positive binding step is often favored to provide extreme levels of purification.
Mimetic Ligand™ adsorbents have intermediate binding specificity; a single adsorbent may be used for the purification of a variety of complementary proteins. Furthermore, a single adsorbent may be suitable for the purification of a number of different proteins. The best adsorbent for a particular need is selected by an initial screen with a PIKSITM test kit. Inexpensive and reusable, PIKSITM modules packed with Mimetic Ligand™ affinity adsorbents enable selection of the ideal adsorbent in a matter of minutes.
Based on our high performance agarose matrices, these screening column sets offer:
- 1ml column volumes
- Packed with most of the ProMetic media currently available
- Convenient to use
- Fast, simple, affordable
- Run with all commercially available instruments, a simple peristaltic pump, or just a syringe
- Easy to scale up
- Compatible around the World
Ready-to-use PIKSI columns are manufactured in biocompatible Tefzel, and include porous top and bottom frits, allowing high flow rates and avoiding "dry out" when the columns are run under gravity.
Ordering Information: PIKSITM Columns:
| Catalog No. | |
|---|---|
| Mimetic™ PIKSITM M Screening Column Kit (10 x 1ml of each Mimetic type) | 1003-00001 |
| Mimetic™ PIKSITM Columns, 5 x 1ml: | |
| Mimetic™ Red 2 A6XL | 1020-00001 |
| Mimetic™ Red 3 A6XL | 1013-00001 |
| Mimetic™ Orange 1 A6XL | 1030-00001 |
| Mimetic™ Orange 2 A6XL | 1040-00001 |
| Mimetic™ Orange 3 A6XL | 1050-00001 |
| Mimetic™ Yellow 1 A6XL | 1060-00001 |
| Mimetic™ Yellow 2 A6XL | 1070-00001 |
| Mimetic™ Green 1 A6XL | 1080-00001 |
| Mimetic™ Blue 1 A6XL | 1090-00001 |
| Mimetic™ Blue 2 A6XL | 1100-00001 |
| Mimetic™ AP A6XL | 1110-00001 |
| Mimeticv Blue SA A6XL | 1120-00001 |
For more information, please contact us.