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Phage Display Library Construction

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Creative Biolabs is pleased to offer customized library construction services under a variety of phage display systems. Our scientists have extensive experience in library construction and are professional in generating different types of phage display libraries, including scaffold libraries, peptide libraries, antibody libraries, cDNA libraries and others.

Phage Display Library Types

Antibody Library (Immune, Naïve, Semi-synthetic, Synthetic)

Creative Biolabs can display the interest antibody in either scFv or Fab format to construct a high-quality antibody library. Due to the smaller size, scFv libraries are genetically more stable than Fab libraries, while Fab libraries lack the tendency to form higher molecular weight species, such as dimers and trimers, which can simplify the selection and characterization. Moreover, both human plasma cells and splenocytes from naïve or immune animals can be accepted as repertoire source for our antibody library construction service. In addition, Creative Biolabs is able to generate fully or semi-synthetic antibody libraries as well.

Antibody Library (Immune, Naïve, Semi-synthetic, Synthetic)

Scaffold Library

Creative Biolabs is able to generate high-quality phage display libraries for a comprehensive list of scaffolds. Our scientists devote to offer customized service to design specific scaffold based on clients’ demands. In addition, to achieve 100% precise mutant and over 1010 diversity, both trimer codon technology and NNK method are used in the library construction resulting in an increased randomicity, which enables the library to be more suitable for sorting and isolating the high-affinity protein or peptide targets.

Peptide  Library (Naïve, Semi-synthetic)

Peptide Library (Naïve, Semi-synthetic)

To form a peptide library, Creative Biolabs can generate constrained structures with target-adapted cross-links for both naïve and semi-synthetic peptides. This structure contributes to maintaining or increasing a series of properties of peptides, such as stability, permeability, binding affinity and resistance to protease activity. Furthermore, for peptide libraries with inserts longer than 8-mer (and up to 30-mer), three nucleotide doping strategies (NNK, Split-mix-split synthesis, and Trimer codon) can be adopted to maximize the amino acid sequences encoded by the inserted DNA sequences.

cDNA Library

According to our advanced phage display platform, Creative Biolabs is capable of displaying entire cDNA products on the surface of appropriate phage system. Compared with conventional cDNA libraries, our high-quality libraries can achieve higher capacity, density, and correct orientation, which further contribute to the investigation of a lot of protein candidates. Moreover, there are various types of cDNA libraries, including standard library, full-length library, normalized library and subtractive library, can be tailored to match our clients’ demands.

Diversity of Semi-synthetic/Synthetic Libraries

In general, Creative Biolabs employs three main approaches to design semi-synthetic or fully synthetic libraries with appropriate diversity. Our scientists normally generate libraries with a diversity of 108, while we are also able to generate libraries over 1010 or follow our clients’ specific requirements.


Phage Display  Systems


Phage Display Systems

Based on the interest protein and specific project objective, Creative Biolabs is able to tailor the most suitable phage display system (M13, T4 or T7) for the library construction.

 


Table 1. Different phage display vehicles and their characteristics (Ebrahimizadeh and Rajabibazl 2014)

  M13 T4 T7
Genome Size 6407 bp 168895 bp 39937 bp
Display Protein All coat proteins, usually pIII and pVIII SOC and HOC gp10B
Display Size >110 kDa on pIII
<10 kDa on pVIII
<710 kDa <132 kDa
Display Density <5 copies on pIII
<2700 copies on pVIII
<810 copies on SOC
<155 copies on HOC
<415 copies
Lifecycle Lysogeny Lytic Lytic

λ Phage

In addition to regular M13, T7 or T4 phage, λ phage is an ideal vehicle to carry the DNA fragments due to its simple structure and flexible DNA. By employing multiple lambda vectors, Creative Biolabs can offer specific genome library construction service to generate qualified genome library with over 95% coverage from either genomic DNA, chromosomes or microbes. In addition, our professional scientists are also available to generate lambda based cDNA library as an alternative service.

As one of the most famous industry leaders who is skilled in advanced phage display technology, Creative Biolabs not only concentrates on developing cutting-edge technology but also be willing to share our state-of-the-art platforms and sufficient expertise with our clients to promote their brilliant studies. It is our guarantee to provide customers with the best services and finest results at Creative Biolabs.

Reference
Ebrahimizadeh, W. and Rajabibazl, M. (2014) 'Bacteriophage vehicles for phage display: biology, mechanism, and application', Current microbiology, 69(2), 109-120.




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