Kansas State University Libraries Kansas State University Libraries
New Prairie Press New Prairie Press
Kansas State University Undergraduate Research Conference Spring 2019
Synthesis and Optimization of Novel Poly-(β-amino ester) Synthesis and Optimization of Novel Poly-( -amino ester) Polymer for Gene-Delivery Polymer for Gene-Delivery
Maria Montes
Follow this and additional works at: https://newprairiepress.org/ksuugradresearch
Part of the Organic Chemistry Commons, and the Polymer Chemistry Commons
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Recommended Citation Recommended Citation Montes, Maria (2019). "Synthesis and Optimization of Novel Poly-(β-amino ester) Polymer for Gene-Delivery," Kansas State University Undergraduate Research Conference. https://newprairiepress.org/ksuugradresearch/2019/posters/70
This Event is brought to you for free and open access by the Conferences at New Prairie Press. It has been accepted for inclusion in Kansas State University Undergraduate Research Conference by an authorized administrator of New Prairie Press. For more information, please contact [email protected].
https://newprairiepress.org/https://newprairiepress.org/ksuugradresearchhttps://newprairiepress.org/ksuugradresearchhttps://newprairiepress.org/ksuugradresearch/2019https://newprairiepress.org/ksuugradresearch?utm_source=newprairiepress.org%2Fksuugradresearch%2F2019%2Fposters%2F70&utm_medium=PDF&utm_campaign=PDFCoverPageshttp://network.bepress.com/hgg/discipline/138?utm_source=newprairiepress.org%2Fksuugradresearch%2F2019%2Fposters%2F70&utm_medium=PDF&utm_campaign=PDFCoverPageshttp://network.bepress.com/hgg/discipline/140?utm_source=newprairiepress.org%2Fksuugradresearch%2F2019%2Fposters%2F70&utm_medium=PDF&utm_campaign=PDFCoverPageshttps://creativecommons.org/licenses/by-nc-nd/3.0/https://creativecommons.org/licenses/by-nc-nd/3.0/https://creativecommons.org/licenses/by-nc-nd/3.0/https://creativecommons.org/licenses/by-nc-nd/3.0/https://newprairiepress.org/ksuugradresearch/2019/posters/70https://newprairiepress.org/ksuugradresearch/2019/posters/70mailto:[email protected]
z
zvvv zzvvv
Synthesis of PBAE-cationic polymer and optimization of
molecular weight for linkage of PBAE polymer to cell
penetrating peptide for further cell transfections.
Bossmann Lab Group,
Bridges to the Future Program,
Developing Scholars Program (DSP),
Johnson Cancer Research Center,
Funding provided by the National
Institutes of Health/National Institute
of General Medical Sciences Award
5R25GM119968-02.
Synthesis and Optimization of Novel Poly-(β-amino ester) Polymer for Gene-
Delivery. Maria Montes-Gonzalez, Obdulia Covarrubias Zambrano, Tej Shrestha, and Stefan H. Bossmann
Kansas State University Manhattan, KS
Department of Chemistry
• Polymer was successfully synthesized and characterized.
•Zeta potential experiments demonstrated optimal conditions to obtain
a positive zeta potential charge, which is ideal for cellular uptake
during a transfection.
•Cell experiments demonstrated successful transfection for 150µM
PBAE incubated for 30 min with GFP plasmid DNA.
•WTAS successfully synthesized and loaded onto cells to determine
cell penetration and toxicity.
1Kim, J.; Kang, Y.; Green, J. Stable and effective poly(ethylene glycol)-
poly(beta-amino ester) nanoparticles for gene delivery. September 1, 2018.
293.301. 2Mangraviti, Antonella, et al. “Polymeric Nanoparticles for Nonviral Gene
Therapy Extend Brain Tumor Survival in Vivo.” ACS Nano, vol. 9, no. 2, 2
Feb. 2015, pp. 1236–1249., doi:10.1021/nn504905q.3Smith, Tyrel T., et al. “In Situ Programming of Leukaemia-Specific T Cells
Using Synthetic DNA Nanocarriers.” Nature Nanotechnology, 17 Apr. 2017,
pp. 1–8., doi:10.1039/NNANO.2017.57.4Wang, Hongwang, et al. “Rationally Designed Peptide Nanosponges for Cell-
Based Cancer Therapy.”
24 hours at 90ºC
+
1,4-butanediol diacrylate 3-amino-1-propanol
Step 1
tris(2-aminoethyl) amine
Polymer Synthesis
+
Gene delivery is one of the most promising methods to treat
multiple diseases through the alteration of a genetic code to
enhance or inhibit gene expression (NIH). The potential for gene
delivery to be used as a treatment for cancerous cells is the
beginning for advanced personalized healthcare. Current methods
for cancer are costly, nonspecific, and come with major side effects
that lowers quality of life in cancer patients. Previous studies have
demonstrated Poly-(β-amino ester) (PBAE) to be biodegradable,
non-toxic, and capable to deliver payloads in a targeted manner.
For this study, PBAE was synthesized and characterized by
Nuclear Magnetic Resonance, Dynamic Light Scattering, and Zeta-
Potential. Cell experiments were conducted to determine
transfection efficiencies. Also, conjugation of PBAE with a cell
nucleus penetrating peptide (WTAS) will be investigated to
develop a more effective gene delivery system, composed of two
nanocarriers. Specifically, focus is emphasized on the advancement
of non-viral gene delivery systems that can have a general
application towards resolving current barriers in gene therapy.
WTAS Live Confocal StudiesStep 2
• Non-toxic,
• Cell penetration to cell nuclei within a couple minutes after exposure.
PBAE Polymer : WTAS : EDC : DMAP
1 : 4 : 4 : 4
2 hours at r.t.
Intermediate 1
50 µM WTAS loaded into B16F10 cells
Peptide Synthesis
(W)TAS : PLKWPGKKKKGKPGKRKEQEKKKRRTR
Fmoc
Fmoc
Fmoc
Positive zeta potential charge is ideal for
cellular uptake through endocytosis, which
was obtained only these PBAE concentrations
and incubation time with GFP plasmid DNA
Linking PBAE to WTAS
T=2 hrsT=0 hrs
Acknowledgements
Abstract
Objective
Transfection experiments Conclusion & Next Steps
References
Poly(β-amino ester)
THF
Incubation of PBAE with DNA at various concentrations and time
Cell transfection experiments in Neural stem cells (NSC)
150µM PBAE (1.05:1) incubated with 500 ng pGFP DNA for 30 min
GFP fluorescent
cells transfected
Non-fluorescent
cells
Merged images
PBAE polymer WTAS C-terminal side
Synthesis and Optimization of Novel Poly-(β-amino ester) Polymer for Gene-DeliveryRecommended Citation
PowerPoint Presentation
Top Related