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The Seamless Construction of Phage Display Libraries

As a long-term expert in phage display technology, Creative Biolabs owns lots of scientists who are proficient in Phage Display Library Construction. By using type IIS restriction enzymes, Creative Biolabs developed an advanced method for the seamless construction of phage display libraries, including phage display Fab/scFv/ VHH library, phage display peptide library, phage display protein variant library, etc. The method is faster, simpler and more reproducible than the two classical procedures, including splicing-by-overlap extension PCR (SOE-PCR) or restriction cloning. Moreover, this strategy could also be applied to the reformatting of antibodies across multiple expression systems.

The Seamless Construction of Phage Display Libraries

For the construction of phage display Fab/scFv libraries, it requires the cloning of variable domains from both the light and the heavy chain of antibodies. In the early days of phage display, libraries construction involved separate cloning of light chains and heavy chains, requiring twice the amount of work compared to production of a single chain display. A simple solution to this problem is to use two plasmids to clone each chain separately. Although this technique has been shown to generate very large libraries, it is still not popular. Currently, most Fab libraries are still built on a single phagemid following well-tested restriction enzyme-based strategies such as those developed for pComb3H or pHEN2, and many others. SOE-PCR is an attractive and popular alternative for antibody library construction. The advantages of SOE-PCR include single step assembly of the variable heavy chain (VH) and light chain (VL), the absence of restriction site-induced mutations and one-step cloning of the final construct into the receiving vector. Despite the advent of powerful strategies, such as SOE-PCR and restriction cloning, obtaining high-quality libraries with excellent coverage is still challenging. These methods require multiple and complex PCR reactions which do not always produce large quantities of properly assembled double-stranded material. For the construction of phage display peptide library or phage display protein variant library, the process is a little bit easier, but the same problem persists.

Creative Biolabs has explored an alternative strategy for the seamless construction of phage display libraries by using type IIS restriction enzymes. Type IIS restriction enzymes have non-palindromic asymmetric recognition sites and cut outside of their recognition sequence, they allow unidirectional cloning with deletion of the recognition site during subsequent ligation. Our scientists combine multiple cloning reactions in a single tube, where the seamless ligation carries on. Then, the validation of library will be conducted by our advanced Magic™ platform.

Key advantages of the seamless construction of phage display libraries including but not limited to:

As an industry leader specialized in custom services, Creative Biolabs provides a series of services in phage display libraries construction, include phage display antibody library, phage display peptide library, phage display protein variant library, etc. We are proud to introduce our cutting-edge technology in the seamless construction of phage display library to all our global customers. We are confident in offering the best service at the most competitive cost. Please feel free to inquire us for more information and a detailed quotation.

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