Along with extensive experience in providing custom scaffold protein library construction services, Creative Biolabs is now able to offer the novel Alphabodies library construction service for our customers all over the world. Based on our proprietary Hi-Affi™ platform, our scientists are confident in tailoring the specific protocols to provide 100% precise mutant library with the expected size of 1010.
Introduction of Alphabodies
In recent years, protein scaffolds have the potential to replace antibodies for various biomedical and biotechnological applications. Now we have developed the affinity-matured Alphabodies which featuring a single-chain antiparallel triple-helix coiled-coil fold. The design and development of Alphabodies as protein scaffolds are not present in nature but with unique physicochemical and structure-function properties. The Alphabody is the ideal drug scaffold to mimic the α-helical domains from disease-causing proteins and therefore inhibit the interactions with other proteins. Moreover, the Alphabody has the ability to pass through the cell membrane and target to the intracellular proteins thanks to the specific positioning of charged and hydrophobic residues.
Construction of Alphabodies Library
In general, an Alphabody is designed to feature threeα-helices composed of four heptad repeats for each one. These three α-helices are connected with two linkers (L1 and L2). So the full amino-acid sequence can be defined as N-HRS1-L1-HRS2-L2-HRS3-C. As we have got the idea of the sequences of heptad repeat HRSx and Lx, it would be possible to build the Alphabody library expediently.
Fig.1 Schematic overview of the Alphabody platform. (Desmet, J.; et al 2014)
In order to generate the target-specific binders, we are able to build the Alphabodies library according to residues randomization. The cDNA of Alphabodies are cloned as the template. After a series of PCR, the genes with the position-specific randomization (NNK) are fused with the pIII gene of bacteriophage M13 and we can get the custom Alphabodies library. According to further biopanning approach, specific Alphabody scaffolds can be selected to target the diverse binding epitopes.
The rationale of the Alphabody scaffold development
Based on our extensive experience and advanced phage display platform, Creative Biolabs now can provide the unique Alphabodies library construction service to meet the specific needs of our clients. If you are interested in our service, please do not hesitate to inquire us for more details.