07 Mar CLICK´N CUT Affinity resin
CLICK’N CUT™ is a universal and robust tool to obtain tag-less proteins in a one-step process based on intein affinity. It is suitable for every step of product and process development: R&D, pre-clinical, clinical and industrial manufacturing.
Inteins are self-cleaving protein domains that can be used in protein purification techniques to selectively isolate a target protein of interest. One way to do this is by using an intein affinity resin, which is a type of chromatography resin that specifically binds to intein-tagged proteins.
The use of intein affinity resin is a popular technique in protein purification because it provides high selectivity, high yield, and high purity of the target protein. Here are some key points to keep in mind when working with intein affinity resin for protein purifications:
1.Choosing the right intein: The choice of intein depends on the specific requirements of the experiment. CLICK´N CUT is a universal tool with full protein compatibility, applicable to any experimental condition
2.Constructing the intein-tagged protein: The target protein of interest is fused to the intein domain using recombinant DNA technology. The intein can be placed at either the N- or C-terminus of the target protein, depending on the specific requirements of the experiment. Once the fusion protein is expressed and purified, the intein can be used to isolate the target protein
3.Purifying the intein-tagged protein: The purified fusion protein is applied to the intein affinity resin, which specifically binds to the intein domain. After washing away non-specifically bound proteins, the target protein is released from the resin by triggering intein-mediated self-cleavage. The released target protein can then be further purified by additional chromatographic steps.
4.Optimizing the purification conditions: The purification conditions, such as pH, salt concentration, and buffer composition, can affect the binding and release of the target protein from the intein affinity resin. Therefore, it is important to optimize these conditions to maximize the yield and purity of the target protein.
In summary, intein affinity resin is a powerful tool for protein purification that provides high selectivity, yield, and purity. By carefully choosing the right intein, constructing the intein-tagged protein, preparing the resin, and optimizing the purification conditions, researchers can obtain highly pure and functional target proteins for downstream applications.