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Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
Published on: January 7, 2019
High-throughput identification of putative receptors for cancer-binding peptides using biopanning and microarray
Daniel J Ferraro1, Sandeep R Bhave, Rama P Kotipatruni
1Department of Radiation Oncology, Mallinckrodt Institute of Radiology, and Siteman Cancer Center, Washington University School of Medicine, 4511 Forest Park, Saint Louis, MO 63110, USA.
Integrative Biology : Quantitative Biosciences From Nano to Macro
|November 14, 2012
Summary
Reverse biopanning is a new high-throughput method to identify cancer-targeting peptide receptors. This approach aids in selecting peptides for drug development by revealing their specific protein interactions, like TIP-1.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Phage-display peptide biopanning identifies cancer-targeting peptides but requires receptor identification for mechanism validation and optimization.
- Current receptor identification methods can be slow, and some peptides bind non-specific proteins, hindering drug development.
- Identifying specific peptide receptors is crucial for understanding cancer-targeting peptide mechanisms and improving drug design.
Purpose of the Study:
- To introduce a high-throughput method, termed "reverse biopanning," for parallel screening of numerous peptides to identify their receptors.
- To demonstrate the utility of reverse biopanning by screening previously identified cancer-binding peptides.
- To enable selection of peptides for further development based on identified receptor interactions.
Main Methods:
- Developed and applied a high-throughput "reverse biopanning" technique for parallel peptide receptor screening.
- Screened a library of 39 cancer-specific peptides using the reverse biopanning method.
- Employed computational modeling and electrophoretic mobility shift assay (EMSA) for validation of peptide-receptor binding.
Main Results:
- Reverse biopanning identified two peptides, RKFLMTTRYSRV and KTAKKNVFFCSV, as candidate ligands for tumor-expressed protein 1 (TIP-1).
- Computational modeling indicated strong binding affinities for selected peptides to TIP-1, comparable to a known TIP-1 binder.
- In vitro EMSA confirmed robust binding of RKFLMTTRYSRV and its truncated form TTRYSRV to TIP-1.
Conclusions:
- Reverse biopanning is an effective high-throughput method for identifying peptide receptors.
- This method facilitates the selection of peptides for drug development based on specific receptor interactions.
- The identified peptide-TIP-1 interactions offer potential for targeted cancer therapies.
