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Matches generated from the mRNA display library need to be converted into lead compounds for further development into therapeutics. CD BioSciences offers mRNA display technology to support the early discovery of peptide drugs. Our services encompass all stages of the early drug discovery process.
Fig 1. General scheme for peptide selection by mRNA display.
Identifying hits for further optimization is critical in any drug discovery effort. For peptide therapeutics, technologies like mRNA display have become powerful tools to discover these new hit ligands against specific targets. To handle the large datasets generated, various methods are employed for analysis, validation and optimization, including hit prioritization, peptide synthesis, and biochemical and biophysical assays.
Fig 2. Workflow of peptide hit identification, prioritisation, and validation. (Yanyan, et al., 2024)
At CD BioSciences, following affinity selection, library members that remain bound to the immobilized targets undergo PCR amplification. This step enriches the mRNA display library for members with higher affinity to the targets. The amplified products are then used for further analysis and validation experiments.
Alternatively, the cDNA of the binding peptides is amplified using PCR and used as a template for the DNA library. This process is repeated through several rounds of mRNA display library screening to enrich peptides with high affinity for the target protein.
The cDNA of the binding peptides is amplified by PCR and sequenced to determine their structure. We use high-throughput sequencing platforms to perform NGS on each round of the DNA library. Sequence data is collected and peptide coding sequences are extracted using Perl scripts. These sequences are further analyzed with FastAptamer software to count and rank unique sequences based on their read numbers.
Peptide Synthesis
Peptides are synthesized using both solid-phase peptide synthesis and peptide biosynthesis. The current mainstream process involves anchoring amino acid units to a solid support with protecting groups, followed by coupling, deprotection, and washing steps. This sequence is repeated to add each amino acid unit until the target peptide is obtained.
Validation and Characterization
The synthesized peptides are purified, analyzed, and validated. Using SPR, Kd values are measured to monitor molecular interactions in real-time and determine the binding affinity of the screened peptides to their ligands. Peptides with strong binding affinity are selected for resynthesis, purification, Kd determination, and subsequent structural optimization.
CD BioSciences' expertise in mRNA display technology, coupled with our robust analysis and optimization processes, positions us at the forefront of peptide drug discovery and development. Contact us for more information about how our innovative mRNA display technology can accelerate your peptide drug discovery efforts and enhance your research outcomes.
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