MTI-2 Library Construction

Creative Biolabs provides professional services for MTI-2 scaffold library construction. With the advanced Hi-Affi™ phage display platform, our top scientists offer libraries with high quality, efficiency and diversity to meet our customers’ specific requirements and facilitate their research and project development.

The mustard trypsin inhibitor 2 (MTI-2), identified in seeds of Cruciferae mustard (Sinapis alba), is a member of Kunitz domain inhibitor scaffolds. Under natural conditions, it is involved in defensive function of plants, which has been shown toxicity for lepidopteran insects. The expression of MTI-2 can be induced in wounded leaves, which may provide a potential tool to study the adaptations of digestive proteases in pest insects. In addition, MTI-2 is a class of trypsin inhibitors that only found in Brassicaceae so far, which do not show any structural homology with other plant proteinase inhibitors. According to these features, MTI-2 is considered as an interesting starting molecule for engineering inhibitory properties.

MTI-2 scaffolds are alternative sources to develop inhibitor with novel specificity. According to structure homology of Kunitz-type protease inhibitors, which is a group of small, irregular a/b-proteins, MTI-2 has been found as a potent, heat-stable polypeptide with 63 amino acids. It is rich in cysteine and glycine residues, presumably eight cysteines form four disulfide bridges to stabilize its secondary construction elements. Naturally MTI-2 has exhibited slow but tight-binding capacity, reversible inhibitors of trypsin, while novel specificity can be select from MTI-2 variants libraries. A typical instance is Chy8, a recombinant chymotrypsin inhibitor selected from MTI-2 phage display library. Therefore, with appropriate mutation sites and library construction technology, MTI-2 is available to be used as scaffold for novel protease targets design.

Scaffold libraries of Creative Biolabs are generated through our proprietary Hi-Affi™ phage display platform, which is based on the phage display technology. The library of target is fused to bacteriophage coat proteins and thereby displayed on the phage surface. Compared with the traditional phage display method, in which the library diversity is insufficient to screen for a high affinity binder for the target, Hi-Affi™ phage display platform, with developed trimer codon technology and NNK method, can introduce more random mutation to expand the range of the library diversity. According to this technology, our scientists have obtained 100% precise mutant library construction with over 1010 diversity.

Creative Biolabs has years of research and development experience in the field of scaffold library construction. Our scientists have successfully generated about 55 kinds of scaffold libraries for clients all over the world by now. It is our guarantee to provide each customer with desired high quality library that fits all individual specifications.

Fig.1 Kunitz type trypsin inhibitor complex with porcine trypsin. (PDB ID: 4AN7)

All listed customized services & products are for research use only, not intended for pharmaceutical, diagnostic, therapeutic or any in vivo human use.

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