mRNA Display Services
Rx Biosciences specializes in constructing and screening mRNA display libraries. These libraries consist of diverse peptide sequences, cDNAs, and antibodies, all designed with specific elements such as sites for T7 RNA polymerase, ribosome binding, actinomycin/puromycin binding, and the RNA stability motifs.
The process begins with in vitro transcription, where the mRNA is synthesized and covalently attached to puromycin. This attachment allows for the mRNA and the protein translated from it to be linked together. The translation itself is carried out in vitro using either E. coli or eukaryotic cell lysates, along with tRNA, buffer, and high concentrations of Mg++.
Once the polypeptide, mRNA, and puromycin complex are formed, it is purified and mixed with the target of interest. Through several rounds of screening, the target-peptide-mRNA-ribosome quaternary complexes are selectively isolated and enriched. The RNA from these complexes is then isolated, reverse transcribed, and cloned into a vector. The clones are subsequently sequenced and translated to determine the amino acid sequence of the ligands.
The ribosome display technique employed by Rx Biosciences allows for the screening of peptide, protein, and antibody libraries with an exceptional diversity of up to 10^14-15. This significantly increases the likelihood of identifying high-affinity binders against a wide range of targets.
Service Highlights
- We create and screen libraries with >10^14-15 diversity.
- Antibody library from immunized animals is prepared and screened.
- Peptide library of different lengths (mini-proteins and small peptides) are prepared and screened.
- cDNA library preparation for mRNA display platform is offered.
- We prepare libraries that incorporate unusual amino acids and circular peptides.
- Finally, identified clones get validated by FACS, Elisa and Biacore.
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