Phage Display Services
Rx Biosciences offers high-quality custom phage display services tailored for biological research and drug discovery initiatives. Our comprehensive approach encompasses library construction and screening, utilizing advanced molecular biology techniques.
We begin by amplifying the antibody repertoire from peripheral blood mononuclear cells (PBMCs) and employing high- throughput (HTP) synthesis of antibody genes. This process includes the randomization of complementarity-determining regions (CDRs) within antibody scaffold sequences, as well as in silico design and de novo synthesis of mini-protein genes. These elements are then cloned in conjunction with phage coat proteins, resulting in libraries that display a diverse array of protein at the surfaces of phages, such as M13 and T7. This surface display allows for effective interaction with target molecules in the
surrounding environment.
To isolate proteins that specifically bind to our target of interest, we employ a binding affinity-based technique known as panning. This selection process typically involves a phage library containing billions of unique displayed proteins. Following the selection phase, we identify the proteins presented by the selected phages through phage amplification and subsequent DNA sequencing.
At Rx Biosciences, we are committed to advancing your research and drug discovery projects with our innovative phage display services.
Schematic diagram showing Phage Display Library Construction and screening.
Our Approach
At Rx Biosciences, we employ a comprehensive strategy to screen both off-the-shelf and custom-prepared phage display libraries against client-supplied targets. Our process begins with the expression of antibodies on the surface of cells, allowing them to bind to the antigen under progressively stringent conditions. After conducting approximately three to four rounds of screening, we isolate individual clones and express them as proteins. The binding characteristics of the isolated single-chain variable fragments (scFvs) are meticulously analyzed using a range of techniques, including ELISA, DNA sequencing, fluorescence-activated cell sorting (FACS), and surface plasmon resonance (SPR). By integrating phage display and yeast display technologies in our combinatorial library screening, we enhance the efficiency of identifying high-affinity binding proteins, ensuring optimal results for our clients.
Service Highlights
- Construction of phage display libraries, including cDNA, peptide, scFv, Fab, and vHH antibodies, with an impressive diversity range of 1×10^10 to 1×10^12.
- Development of comprehensive antibody libraries derived from human, mouse, rabbit, chicken, and llama sources.
- Creation of custom peptide libraries utilizing a modified T7 select vector specifically designed for Phage Immuno-Precipitation (PhIP) experiments.
- Screening of Ph.D. 7, Ph.D. 12 and Ph.D. C7C libraries from NEB
- Rigorous screening of various phage display libraries to identify high-affinity binders.
- Expression and purification of scFv and Fab antibodies in bacterial systems, ensuring high yield and quality.
- Conversion of scFv and Fab antibodies into humanized IgGs, followed by expression in mammalian cells for enhanced functionality.
- Validation of binding affinities using advanced techniques such as ELISA, Biacore, and flow cytometry systems, ensuring precise and reliable results.
These services are designed to support cutting-edge research and development in antibody engineering and therapeutic applications.
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