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July 18, 2013 -- Together with Creative BioLabs, Creative Diagnostics offers a unique bacterial display technology, Hi-affinityTM, for the development of human antibodies with pM affinity, the highest affinity ever enabled by an in vitro antibody production technology.
Bacterial display (or bacteria display or bacterial surface display) is a protein engineering technique widely used for in vitro protein evolution. Libraries of polypeptides displayed on the surface of bacteria are screened using flow cytometry or iterative selection procedures (biopanning). Hi-affinityTM is based on a proprietary synthetic bacterial display human antibody library and a unique selection process natural to E. coli. This bacterial surface display technology enables the rapid isolation of target-specific antibodies without the labor intensive screening common to other recombinant and non-recombinant antibody production methods, resulting in unique single-chain variable fragment (scFV) antibodies that have both high specificity and extremely high affinity [Kd up to10-12] for the target antigen.
Hi-affinityTM human scFv antibody library is created by combining a highly diverse collection of synthetically-constructed randomized CDR sequences that are further diversified by random lengths using a unique proprietary technique. The large library has been specifically optimized to eliminate unwanted stop codons and aggregation-prone sequences. scFv molecules are first expressed in E. coli cytoplasm and then translocated and anchored into the bacterial plasma membrane. In the end, binder panels are selected by FACS or panning. Furthermore, multiple rounds of affinity maturation during library screening are incorporated through error prone PCR mutagenesis either directed at CDR or flanking sequences and selection by varying antigen dose.
The Hi-affinityTM platform utilizes a unique antibody selection process that relies on the natural twin-arginine translocation (Tat) system in E. coli. The major limitation challenging conventional phage display technologies is their dependence on the Sec translocation pathway. The Sec pathway transports proteins from the cytoplasm to the periplasm in an unfolded state; consequently, proteins that require a cytoplasmic environment and/or cytoplasmic components for folding, or reach their native state before they interact with the Sec proteins, are not compatible with the Sec pathway. Hi-affinityTM platform overcomes this limitation by exploiting the properties of the Tat pathway which only exports folded proteins that have already attained their native conformation in the cytoplasm.
Hi-affinityTM derived antibodies are intended for research, diagnostic and therapeutic use. The customer owns the exclusive rights to the antibodies including the antibody CDR sequence and any information generated by the customer through using the antibodies.
The featured benefit of this technology is its capacity of generating human antibodies with exceptionally high Affinity.
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