Phagemid Display System pSKAN - Construction of Libraries with Huge Molecular Diversity. The pSKAN Phagemid Display System allows selection and production of novel proteins with the desired affinities to a given epitope. pSKAN is a combinatorial phagemid library expressed in E. coli: The phage coat protein pIII is fused to human pancreatic secretory trypsin inhibitor protein (hPSTI), which contains a short stretch of hypervariable amino acids. This loop is exposed and displays about 3 x 107 randomly recombined amino acid variants. They can be screened in a "panning assay" for specific binding to a target molecule of interest. Variants with optimal affinity are selected and propagated in E. coli. To determine the sequence of a selected hypervariable loop, two sequencing primers are available. MoBiTec provides three pSKAN libraries which differ in the extent of their hypervariable loop: HyA, HyB and HyC. Antibodies against pIII and hPSTI can be obtained for Western blots. For the production of soluble hPSTI protein with the desired binding properties, the vector pMAMPF3-PSTI4 can be used. To construct your own peptide library, pSKAN8 vector DNA is available. Features: • Easy-to-handle • Fast (results in less than 3 weeks!) • Low cost • Lab animals or tissue cultures become superfluous • Direct physical link between genotype and phenotype: DNA codes for addressable tag on phage surface • Construction of libraries with enormous molecular diversity • pSKAN phagemid targets small, recessed epitopes • Stability of libraries due to strong repression of lambda-pL-promoter. Applications: • High-throughput screening of protein-protein interactions • Detection of proteins with specific binding properties (high affinity binders) • Protease inhibitor screening • Development of affinity matrices • Ligand screening • Development of pharmaceuticals and diagnostics • Detection of allergens • Of special interest: 3-D structure-based protein design |
