Ribosome Display

Overview of Ribosome Display

Ribosome display is an in vitro protein evolution technology used to identify proteins that strongly bind to targets. Other methods similar to this technique include phage display, mRNA display, and yeast display. In addition to being used to study protein-protein interactions that occur in daily metabolic processes, ribosome display also allows scientists to identify proteins and antigens present in certain diseases, develop new drugs to block pathways in the body, and understand how proteins interact in normal biological systems.

Principle of Ribosome Display

The principle behind ribosome display is the access to large databases of DNA sequences encoding proteins. These sequences are transcribed and translated in vitro, but the stop codons of each gene are removed and replaced with spacer sequences, small segments of non-coding DNA. This allows the ribosome to translate the gene into a protein product while keeping all components together. Magnesium ions and low temperature help to maintain stability of RNA, ribosomes, and proteins. Affinity chromatography is then used to bind target proteins to beads in the column, followed by running samples from transcription and translation of the DNA library. If any proteins in the DNA sequence have affinity for the target protein, they will be bound and retained in the column. They are then eluted and the mRNA is reverse transcribed to cDNA and amplified using PCR. This cDNA can undergo mutagenesis. Ribosome display is then carried out on this modified DNA with greater selective pressure to produce proteins with higher affinity for the target protein, leading to strong binding selection.

Fig. 1 Principle of in vitro ribosome display.¹

Applications of Ribosome Display

• Drug Design: Proteins that strongly bind to receptors and other proteins can be designed through ribosome display to develop inhibitors or stimulators that can affect many pathways and prevent diseases. For example, peptides discovered on the surface of Staphylococcus aureus have been identified through ribosome display, providing a new avenue for antibiotic development.
• Antibody Optimization: Ribosome display is an important mechanism for protein optimization that can improve the binding strength and stability of antibodies. Antibodies with 40 times stronger affinity than the original can be produced. This can also help further understanding of the interaction between antibodies and cancer biomarkers in cells.
• Nanotechnology: Nanoparticles, as carriers of drugs and other biomolecules, require target binding ligands, which can also be designed through ribosome display.

The main advantage of ribosome display is its ability to link genotype and phenotype by combining mRNA and its encoded protein in the same complex. This allows for easy amplification using PCR once the protein has been identified. Additionally, ribosome display does not rely on the ability of cells to be transformed like other techniques do. Moreover, this method also allows for simple mutations to be made to selected DNA sequences without the need to modify the original library. Creative Biolabs specializes in ribosome research and provides a wide range of services to global clients, including but not limited to Ribosome Analysis Services and Ribosomal Marker Antibody Development Services. If you are interested in our services, please don't hesitate to contact us for more information.

Reference

1. Kunamneni, Adinarayana, et al. "Ribosome display technology: applications in disease diagnosis and control." Antibodies 9.3 (2020): 28.