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Phage Display

PSKAN - pSKAN Phagemid Display System


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The pSKAN Phagemid Display System is a fast and easy to handle alternative to antibodies or immunoglobulin domain presenting phagemids. Since the pSKAN phagemid displays a small hypervariable amino acid loop, more than 3 x 107 variants can be tested for the specific binding to a target molecule of interest. The variants with the optimal affinity can be selected and propagated without having to use animals or tissue cultures. The sequence of the binding domain can be determined easily by DNA sequencing. In comparison to antibodies, the pSKAN System is able to target very small epitopes, since it searches for binding sites with "only a fingertip" instead of "two whole hands" like antibodies. Within a peroid of only three weeks, the scientist is able to receive the target molecule of interest.

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.

CONTENTS: The system consists of 3 libraries HyA, HyB, and HyC, including the host strain. Sequencing primers, detection antibodies, and additional DNA vectors are available separately