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Monobody/Adnectin/FN3 Scaffold Library Construction

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Creative Biolabs offers custom scaffold library construction services for monobody. Our innovative technology enables generation of scaffold libraries with desired diversity and high controllability.

Monobodies are recognized as a kind of synthetic binding proteins that developed from fibronectin type III domain (FN3). FN3, as a kind of antibody mimics scaffold, is a simple and robust alternative to antibodies for creating target-binding proteins. FN3 is evolutionary conserved which can be found in a wide variety of animal species and more sporadically in yeasts, plants and bacteria. Typical monobody is around 94 amino acids (about 10 kDa) Ig-like protein, which got similar size with single domain antibody (12-15 kDa) and much smaller size than common antibodies (150-160 kDa). It consists of a small, monomeric natural β-sandwich fold possesses seven β-strands and 2-3 loops connecting the strands in a pairwise fashion at the end of each β-sheet. Notably, monobody is one of the rare members of the Ig superfamily devoid of disulphide bonds. In contrast to antibodies, the fold does not rely on disulfide bonds, yet still displays high thermodynamic stability. Since disulfide bonds do not form in reducing intracellular environments, monobody can be a good choice in such circumstances.

According to the specific properties, diversified monobody library is an ideal choice for the development of binding proteins for various targets. A typical example of this case is firstly demonstrated by monobody generated from combinatorial libraries of human tenth fibronectin FN3 domain (10FN3 domain). Currently, combining with directed evolution technologies, a great deal of monobodies with high affinity and specificity have been reported to bind their respective targets. For instance, high affinity and isoform-specific monobodies were developed as inhibitors of small ubiquitin-related modifier (SUMO) family proteins for their mechanistic and cellular investigations. As well as human anti-PRLR monobody was developed as an innovative treatment for gynecologic cancer. Therefore, it seems reliable to treat monobodies as alternative scaffolds that aim to overcome shortcomings of natural antibody molecules. To achieve this, Creative Biolabs can generate the libraries and screen the stable, functional and specific monobodies to meet our clients’ research objectives

Scientists from Creative Biolabs have applied their analysis and design experience to develop the Hi-Affi™ phage display platform. This robust platform provides an easy and reliable way to build up scaffold libraries and to select potential candidates for use in R&D applications, drug discovery and the development of diagnostic kits. In general, we can generate high affinity FN3 phage display library with 100% precise mutant and over 1010 diversity. In addition, it is also available for Creative Biolabs to deliver successful FN3 library based on different techniques, such as ribosome display and mRNA display.

Our proprietary Hi-Affi™ platform features:

• High affinity
• Controllable mutagenesis
• Applicable to high-throughput methods
• High diversity
• Fast turnaround time

Creative Biolabs is the right company who can speed up your research and project development. Our devoted scientists are pleased to guarantee our customers high quality scaffold libraries with desired diversity and operability. We have established long term relationship with most of our customers, and the convenience and flexibility is within the objectives we are desperate to achieve when serving them.

Fig. 1 The tenth fibronectin type III domain. (PDB ID: 1TTG)

Fig. 2 Monobody in complex with the SH2 domain. (Jost and Plückthun 2014)

Reference
Jost, C. and Plückthun, A. (2014) 'Engineered proteins with desired specificity: DARPins, other alternative scaffolds and bispecific IgGs', Current opinion in structural biology, 27, 102-112.




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