Ribosome Display

Ribosome display library construction plays a crucial role in identifying high-affinity peptides, cDNAs, and antibodies. At Rx Biosciences, we specialize in constructing and screening ribosome display libraries to support efficient in vitro selection of binding molecules.

We begin by generating a diverse library of peptides, cDNAs, and antibody variants. Each sequence includes essential components such as a T7 RNA polymerase promoter, a ribosome binding site, actinomycin/puromycin binding sequences, and RNA stability elements. These features ensure efficient transcription and stable complex formation.

After preparing the library, we perform in vitro transcription to synthesize RNA transcripts. Next, we carry out translation using E. coli cell lysates, tRNA, appropriate buffers, and high concentrations of Mg²⁺ ions. To promote quaternary complex formation, we maintain the process at 4°C. These complexes consist of the ligand, its mRNA, and the ribosome, all bound together.

Through three to four iterative screening rounds, we isolate and enrich complexes that show strong binding affinity. Following each round, we extract RNA from the selected complexes. Then, we reverse transcribe the RNA into cDNA and clone it into suitable vectors.

To identify the selected ligands, we sequence the cloned DNA and translate it into corresponding amino acid sequences. This final step reveals the structure and binding potential of the molecules we’ve selected.

Thanks to this systematic approach, Rx Biosciences helps researchers efficiently discover novel ligands for therapeutic, diagnostic, and research applications. Our expertise in ribosome display library construction ensures accurate, high-throughput screening with reliable results.