EGFR Protein Inhibitor & Nanocarrier Design
Designed a novel therapeutic protein inhibitor targeting the ligand-binding region of the Epidermal Growth Factor Receptor (EGFR) to suppress cancer cell proliferation. Developed a comprehensive manufacturing pipeline using bacterial expression vectors and designed a lipid nanoparticle (LNP) delivery system for targeted cancer therapy.
Technical Skills Demonstrated
Rational Protein Design
- De novo peptide design (20 residues) targeting EGFR active site
- Complementary amino acid selection (Ser/Thr, Gln/Asn)
- Structure prediction utilizing AlphaFold/ColabFold
- Ligand-binding domain analysis (PDB: 3NJP)
Genetic Engineering & Cloning
- Reverse translation of peptide to DNA sequence
- pUC19 expression vector design with AgeI/NcoI sites
- Primer design and melting temperature optimization (~36-38°C)
- PCR protocol development for gene amplification
Nanocarrier Formulation
- Lipid Nanoparticle (LNP) delivery system design
- Hydrophobic core encapsulation strategy
- Active targeting via GE11 peptide ligand integration
- Particle sizing for small unilamellar vesicles (20-100 nm)
Structural Bioinformatics
- Hydropathy plot analysis of transmembrane domains
- Visual Molecular Dynamics (VMD) analysis
- Protein hydrophobicity and solubility assessment]
- Beta-strand formation analysis
Key Project Outcomes
20 AA
Inhibitor Length
Designed peptide (DNTAVCMTDYACNLGTNCNC) to block EGF binding
GE11
Targeting Ligand
Integrated high-affinity ligand to enhance cancer cell specificity
pUC19
Expression Vector
Circular plasmid designed for E. coli manufacturing
Engineering Analysis: The project successfully established a theoretical framework for inhibiting EGFR-driven tumors. By designing a short 20-residue peptide utilizing complementary amino acids (Ser, Gln, Leu), the inhibitor aims to compete with natural EGF ligands. The manufacturing strategy utilizes a pUC19 vector with specific restriction sites (AgeI, NcoI) for precise gene insertion. For delivery, a hydrophobic core LNP formulation was selected to protect the peptide, decorated with GE11 ligands to ensure high-affinity targeting of the transmembrane-juxtamembrane region of cancer cells.
Complete Final Project
Detailed protein structure analysis, plasmid maps, and LNP formulation protocols