mRNA Display-based Protein Screening

mRNA display is a cell-free display technology that can select proteins from a large recombinant DNA library in vitro. It also allows mutations to be continuously introduced into the selected DNA library through PCR-based mutations in each cycle, thus enabling the selection of protein variants with improved consistency, specificity and stability, thus providing a powerful "protein evolution" tool for optimizing therapeutic and diagnostic proteins. CD BioSciences is committed to applying in vitro display technology to the field of protein research, and has accumulated rich experience in screening and guiding protein evolution over the years, which can provide customers with more flexible, fast and cost-effective protein screening and analysis services.

Introduction of mRNA Display for Protein Screening

Protein screening uses mRNA display technology, which allows a large number of mutants to be examined in a single experiment. The protein displayed by mRNA is a structure of covalent connection between a protein and the mRNA encoding it. The covalent linkage between phenotype (protein) and genotype (RNA) effectively makes the protein directly amplified. This connection enables continuous selection, amplification and optional mutation cycles on protein libraries larger than 1012. The size of this library is much larger than that used by most in vivo and in vitro screening methods, and several orders of magnitude larger than that used by traditional screening techniques. The mRNA display program is completely carried out in vitro, which makes it possible to select under various interesting conditions that are incompatible with traditional in vivo selection methods (salt, pH, additives, etc.). If RNA stability is concerned in some environments, RNA can be replaced by its cDNA. mRNA display has been used to select specific ligands from libraries of proteins, peptides, cyclized peptides and peptides constructed from unnatural amino acids.

Overview scheme of the selection of enzymes for bond-forming reactions by mRNA display.Fig. 1 Overview scheme of the selection of enzymes for bond-forming reactions by mRNA display. (Seelig B, 2011)

Key Steps of mRNA Display for Protein Screening

  1. PCR construction/library generation.
  2. Genotype-phenotype complex is produced through cell-free transcription and translation.
  3. Screening the immobilized target.
  4. In situ RT-PCR recovery.
  5. Regeneration of full-length constructs for subsequent mRNA display cycles.
  6. Cloning, expression and screening of specific protein binding agents.

Our Services

The protein discovery platform built by CD BioSciences based on mRNA display technology can select functional proteins from the library of more than 1012 different mutants in a single test tube in vitro, and can be used for high-throughput screening of proteins and new enzymes targeting cell epitopes, GPCR ligands, enzyme substrates, cell signals and other ligands. We provide one-stop service for protein screening for global customers. From the construction of DNA library to the validation of final target molecules, you only need to provide project information. Our professional team will design and optimize the best solution to accelerate your drug discovery or mechanism research cycle.

Service Features

  • Accurate and fast experiment process, saving time for customers.
  • Professional technical experts accurately control the database building process.
  • The constructed library can screen a large number of different targets.
  • The protein screened has high specificity and sensitivity.

CD BioSciences has many years of project development experience in protein discovery. The protein discovery platform based on mRNA display technology can provide customers with services related to the development and screening of various proteins. If you are interested in our services, please contact us for more details.

Reference

  1. Seelig B. (2011). mRNA display for the selection and evolution of enzymes from in vitro-translated protein libraries. Nature protocols, 6(4), 540–552.
For Research Use Only. Not For Clinical Use.

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