1. Design a biotin-labeled peptide library: overlapping or Alanine scan
2. The peptide library is coated on the avidin-coated 96 well plates.
3. The Peptide library is used to evaluate the binding of each peptide to different cell lines using CyQUANT Cell Proliferation Assays.
4. Flow cytometry: Determine the cellular uptake of selected FITC-labeled peptides.
5. Competitive assay: The cells are incubated with FITC-peptide with or without excess unlabeled peptide (100-fold)
6. Finetune the peptide sequence: structure analysis, cyclization, and D amino acid modification.
7. Find the promising receptor-specific cancer cell binding peptide that can be conjugated directly to a chemotherapeutic drug or to nanoparticles for targeted drug delivery to enhance the efficacy of chemotherapy for cancer treatment.

Reference:

https://www.nature.com/articles/s41598-019-38574-y