Episomal Vectors for Stable Production of Recombinant Proteins and Engineered Antibodies
Abstract
:1. Introduction
2. Materials and Methods
2.1. Vector Construction
2.2. Cloning of Single Proteins (e.g., Srt4M) Plasmids
2.3. Cloning of Antibody Plasmids
2.4. Single Plasmid for Bispecific Production
2.5. Cells, Transfection, and Selection
2.6. Antibody Expression and Protein Purification
2.7. Sortase Signal Peptide Library Construction
2.8. Signal Peptide Library PCR Amplification and Sequencing
2.9. Antibody and Protein–Fc Constructs
2.10. siRNA
2.11. Antibody–siRNA Conjugation
3. Results
3.1. Secreted Expression of Sortase A Δ59 Variant Srt4M in Expi293
3.2. Stable Production of Antibodies with Heavy-Chain Tags
3.3. Utilizing Inteins for Production of a Bispecific Antibody-like Molecule
3.4. Split Inteins for Stable Production of Defined Multi-Tag Antibodies
3.5. Site-Specific DAR1 siRNA–Antibody Conjugate
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Fallahee, I.; Hawiger, D. Episomal Vectors for Stable Production of Recombinant Proteins and Engineered Antibodies. Antibodies 2024, 13, 18. https://doi.org/10.3390/antib13010018
Fallahee I, Hawiger D. Episomal Vectors for Stable Production of Recombinant Proteins and Engineered Antibodies. Antibodies. 2024; 13(1):18. https://doi.org/10.3390/antib13010018
Chicago/Turabian StyleFallahee, Ian, and Daniel Hawiger. 2024. "Episomal Vectors for Stable Production of Recombinant Proteins and Engineered Antibodies" Antibodies 13, no. 1: 18. https://doi.org/10.3390/antib13010018