Product/Service
Affinity Purification System - Profinity eXact Fusion-Tag System
•Datasheet: Profinity eXact Cloning and Expression Kits
•Brochure: Profinity eXact Fusion-Tag System
•Datasheet: Profinity eXact Resin
•Technical Note: Profinity eXact Fusion-Tag System Performs On-Column Cleavage And Yields Pure Native Protein From Lysate In Less Than An Hour
The Profinity eXact fusion-tag system is an affinity tag-based protein purification system that addresses the bottlenecks commonly associated with tag removal in other affinity tag systems. This E. coli-based expression and purification system generates a tag-free, highly purified protein containing its native N-terminal amino acid sequence in a single step — without the addition of protease.
Purification and on-column tag cleavage using the Profinity eXact fusion-tag system. During sample application, immobilized subtilisin protease (ligand) recognizes and binds the affinity-tagged protein. Washing the column removes unbound contaminants. Application of fluoride-containing elution buffer triggers subtilisin to quickly and precisely cleave the tag from the fusion protein after the FKAL cleavage recognition sequence. The tag remains tightly bound to the resin, and a highly purified protein with only its native amino acid sequence is released.
The Profinity eXact fusion-tag system utilizes a modified form of the subtilisin protease, which is immobilized onto a chromatographic support and used to generate pure, tag-free target protein in a single step. The tag in this system is the prodomain (prosignal sequence) of the subtilisin protease, a 75-amino acid sequence that is fused to the N-terminus of a target protein of interest. The mature and prodomain subtilisin protease sequences have been co-engineered to produce a highly specific, high-affinity interaction between the binding partners. Application of elution buffer triggers subtilisin's processing activity, which quickly and precisely cleaves the tag from the fusion protein and releases the purified target protein. At the end of the purification process, the tag remains tightly bound to the resin and contains only its native amino acid sequence. The structure and activity of the native protein is maintained, and the need for additional materials, such as cleavage enzymes and purification resin for postcleavage enzyme removal, is eliminated.
The Profinity eXact fusion-tag system is a family of expression, purification, and analysis products, consisting of expression vectors, competent cells, SOC growth media, loose and prepacked purification resin, and detection reagents.
- Rapid purification and on-column cleavage
- Novel alternative to existing affinity tag and tag removal technologies*
- Reduced operating costs, purification steps, and reagent use
- High purity of tag-free target protein
•Datasheet: Profinity eXact Cloning and Expression Kits
•Brochure: Profinity eXact Fusion-Tag System
•Datasheet: Profinity eXact Resin
•Technical Note: Profinity eXact Fusion-Tag System Performs On-Column Cleavage And Yields Pure Native Protein From Lysate In Less Than An Hour
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