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Knottins Library Construction

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Creative Biolabs offers knottins phage display library construction service to facilitate the rapid production and selection of knottins. Creative Biolabs provides a rapid high-throughput parallel library generation platform Hi-Affi™ that enables the construction of knottins libraries with guaranteed quality and more competitive cost than traditional methods.

Knottins, widely known as cystine-knot miniproteins, have emerged as a structural family. This family features a conserved core of three tightly woven disulfide bonds and highly diverse loop regions, which are anchored to a core of anti-parallel β strands and constrained by this molecular “knot” topology. Knottins are typical example of convergent evolution, in which various family members have involved many species, including fungi, plants and animals, and perform different functions, such as ion channel blockade, antimicrobial activity and protease inhibition.

Knottins possess attractive features to be developed as scaffold. The conserved structure imparts extraordinary thermal and proteolytic stability and high levels of sequence diversity among knottins family members. Based on the high stability, knottins are believed to be nonimmunogenic and their small size has translated into desirable pharmacokinetic and biodistribution properties. Moreover, they are amenable to be produced through chemical synthesis or recombinant expression. Currently, it also available to search standardized information of knottins family members from the KNOTTIN database (http://knottin.cbs.cnrs.fr/). Together these attributes make knottins promising molecular scaffolds for protein engineering and translational applications as therapeutic and diagnostic agents.

To generate high quality scaffold libraries, Creative Biolabs has developed the proprietary Hi-Affi™ phage display platform. On the basis of routine phage display technology, which is an exogenous gene expression method through fusing the target genes to bacteriophage coat proteins then displaying on the phage surfaces to select specific binders, our scientists also integrate trimer codon technology and NNK method to introduce more randomicity and diversity. Ultimately, this novel platform can be used to generate high affinity knottins phage display library with 100% precise mutant and over 1010 diversity. In addition, it is also available for Creative Biolabs to generate knottins libraries in different delivery format such as ribosome display, mRNA display, cell surface display and so on.

Our proprietary Hi-Affi™ platform features:

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

Hi-Affi™-based knottins libraries are compatible with various high-throughput screening and selecting methods such as fluorescence-activated cell sorting. We are also providing knottins libraries constructed by synthetic and recombinant methods, and other services such as measurement of binding affinity of knottins to target proteins.

Creative Biolabs has extensive experience in the research and development of scaffold library construction. Over the past decade, our professional scientists have generated about 55 kinds of scaffold libraries for clients all over the world. We are pleased to offer high quality knottins library with best services to meet our customers’ demands.

Knottins Library Construction




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