Nanobiotechnology, gene therapy, and liquid biopsy
Our major research interests are in polymer engineering of nanomaterials and nanotechnology with applications to Bio-Micro-Electro-Mechanical Systems (BioMEMS). MEMS emerged from IC manufacturing and is gaining applications in biomedical and optical communications fields. In the nano- or micron-size range, surface forces play important roles. We need to re-examine the transport equations as well as constitutive relations. In BioMEMS, we are developing affordable new techniques for micro/nanoarray and micro/nanofluidics biochips, and biosensors, multifunctional nanoparticles for drug and gene delivery, and cell-based drug delivery devices. Major applications are cancer detection and treatment, and cell reprogramming for regenerative medicine.
Professor Lee founded and led the NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices (CANPBD) from 2004 to 2015.
He conducted pioneering research in polymer nanocomposites and foams, combining nanocomposites and supercritical carbon dioxide to produce high-performance, thermal-insulating nanocomposite foams for industrial applications, with the potential to replace solid plastics and conventional foams for significant cost savings in materials and energy.
In the area of bio-micro/nanoengineering and micro/nanofluidics, his innovative combination of polymer-based micro/nanoengineering and nanofluidics led to new materials and devices that have the potential to greatly benefit future medical diagnosis, gene therapy, and drug delivery technologies.
For example, his Tissue Nanotransfection (TNT) innovation was named one of 5 breakthrough medical technologies for 2017 by Forbes magazine, and #1 out of 20 life-changing medical breakthroughs for 2017 by Prevention magazine.
The device painlessly injects genetic code into the skin, transforming cells into any type of cell desired, enabling the body to repair injuries or damaged organs. Details of the study appear in Nature Nanotechnology (August 2017).
His recent work published in Nature Biomedical Engineering (January 2020) on Cellular Nanoporation (CNP) enables the use of engineered exosomes, nanoparticles released by cells, as a universal nucleic-acid carrier for medical applications requiring transcriptional manipulation.
He is no longer accepting graduate students.
- Founder and leader of the NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices (CANPBD) since 2004
- Tissue Nanotransfection (TNT)
- Cellular Nanoporation (CNP)
- 26 patents and patent applications, with 17 patents involving innovative research in polymer nanocomposites and biotechnology successfully licensed to industry
Notable Awards and Honors
- Life Achievement Award from the Society of Advanced Molding Technology
- Honorary Professorship at Peking Union Medical College in Beijing China
- International Award, Society of Plastics Engineers, 2010
- Malcolm E. Pruitt Award, Council of Chemical Research, 2008
- Engineering/Technology Award, Society of Plastics Engineers, 2008
- OSU Distinguished Scholar Award, 2000
Awards Chronologically Listed
2017 Foreign Master Professorship at Dalian University of Technology
2017 OSU, College of Engineering Lumley Interdisciplinary Research Award
2012 OSU, College of Engineering Research Award
2010 International Award, Society of Plastics Engineers
2010 OSU, College of Engineering Lumley Interdisciplinary Research Award
2008 Engineering/Technology Award, Society of Plastics Engineers
2008 Malcolm E. Pruitt Award, Council of Chemical Research
2006 Fellow, American Institute for Medical and Biological Engineering
2005 OSU, College of Engineering Research Award
2002 Finalist, Frank Annunzio Award, Christopher Columbus Fellowship Foundation
2002 OSU, College of Engineering Scott Senior Faculty Award
2002 OSU, College of Engineering Lumley Interdisciplinary Research Award
2001 Fellow, Society of Plastics Engineers
2000 OSU Technology Partnership Alliance Award
2000 OSU Distinguished Scholar Award
2000 OSU, College of Engineering Annual Research Accomplishment Award
1997 East China University of Science and Technology, Honorary Professorship
1996 OSU, College of Engineering Annual Research Accomplishment Award
1995 OSU, College of Engineering Research Award
1991 OSU, College of Engineering Research Award
1989 OSU, College of Engineering Harrison Faculty Award for Excellence in Engineering Education
1988 OSU, College of Engineering Research Award
1987 Central Ohio AIChE Section, Innovation in Chemical Engineering
1986-2011 15 Best Paper Awards in Society of Plastics Engineers, Society of Plastics Industry and American Association of Pharmaceutical Scientists Annual Conferences
1985 OSU, College of Engineering Research Award
1983-86 Amoco Foundation Young Faculty Development Award
1982 Rohm & Haas Young Faculty Award
Biomedical Research Areas
Our major research interest is to design and develop micro- and nano-scale biochips/devices for diagnostic and therapeutic biomedical applications including cancer diagnosis/therapy, infectious diseases and regenerative medicine.
For diagnosis, our current focus is on extracellular vesicles (EVs) based liquid biopsy assay development by using affinity based EV separation to sort and capture specific exosome-like and microvesicle-like EV subpopulations and then using molecular beacons and fluorescence labelled antibodies as probes to measure RNA and protein targets at single EV level on a total internal reflection fluorescence (TIRF) microscope.
For therapeutics, we have extended our nanocarrier strategy from synthetic lipoplex and polyplex nanoparticles to cell secreted therapeutic exosomes (tExos) in recent years. Unlike synthetic nanocarriers, tExos possess low toxicity and low immunogenicity, and can penetrate physiological barriers such as blood-brain barrier (BBB) and solid tumors via transcytosis. Exosomes released from stem cells such as mesenchymal stem cells (MSCs) and induced pluripotent stem cells (iPSCs) also provide anti-inflammation and anti-fibrosis functions. Furthermore, we have developed a unique nanochannel electroporation (NEP) technology which can transfect individual cells with high dose control and cell viability. In addition to direct cell transfection of plasmid DNAs and molecular probes for cell reprogamming, gene editing and living cell interrogation, NEP can also produce a large sclae EVs including exosomes from stimulated cells, which containing targeting peptides and therapeutic mRNAs, microRNAs and shRNAs. This technology has been demonstrated in in vivo tissue nano-transfection (TNT) for tissue repair, and in ex vivo cellular nanoporation (CNP) of tExo generation for transcriptional manipulation.
Non-viral Gene Delivery by Nanochannel Electroporation- The ability to deliver precise amounts of biomolecules and nanofabricated probes into living cells offers tremendous opportunities for biological studies and therapeutic applications. It may also play a key role in the non-viral generation of engineered stem cells and induce pluripotent stem cells with high efficiency and non-carcinogenic properties. Currently-available transfection approaches are heavily dependent upon diffusion- and endocytosis-based mechanisms, which results in highly stochastic transfection. We have overcome this problem by developing a new technology, nanochannel electroporation (NEP) allowing transfection of many small sized and delicate cells with precise control over dose and timing. Cell mortality from NEP is virtually zero. We show dose control effects on a variety of transfection agents such as oligonucleic acids, molecular beacon, quantum dots and efficient delivery of large DNA directly into the nucleus using nanoparticle “bullets.” Dosage controlled delivery to multiple cells is not achievable with any existing techniques. NEP also leads to mass secretion of extracellular vesicles containing functional RNAs from transfected cells.
• P. E. Boukany, A. Morss, W-C Liao, B. Henslee, X. Zhang, B. Yu, X. Wang, Y. Wu, H.C. Jung, L. Li, K. Gao, X. Hu, X. Zhao, O. Hemminger, W. Lu, G. Lafyatis and L.J. Lee, “Nanochannel Electroporation Delivers Precise Amounts of Biomolecules into Living Cells”, Nature Nanotechnology, 6, 747-754 (2011), research highlight in Nature Methods, 8, 996-997 (2011).
• D. Gallego-Perez, L. Chiang, J. Shih, J. Ma, S. Kim, X. Zhao, X. Wang, P. Mao, K.J. Kwak, Y. Wu, L. Wu, G. Lafyatis, D.J. Hansford, I. Nakano, and L.J. Lee, “On-Chip Clonal Analysis of Glioma-Stem-Cell Motility and Therapy Resistance”, Nano Letters, 16(9), 5326-5332 (2016).
• D. Gallego-Perez, D. Pal, S. Ghatak, V. Malkoc, N. Higuita-Castro, S. Gnyawali, L. Chang, W-C Liao3, J. Shi, M. Sinha, K. Singh, E. Steen, A. Sunyecz, R. Stewart, J. Moore, T. Ziebro, R.G. Northcutt, M. Homsy, P. Bertani, W. Lu, S. Roy, S. Khanna, C. Rink, V.B. Sundaresan, J.J. Otero, L.J. Lee and C.K. Sen, "Topical Tissue Nano-transfection Mediates Non-viral Stroma Reprogramming and Rescue", Nature Nanotechnology, :10.1038/nnano.2017.134 (2017).
• Z. Yang, J. Shi, J. Xie, Y. Chen, J. Sun, T. Liu, Y. Zhao, X. Zhao, X. Wang, Y. Ma, V. Malkoc, C-L Chiang, Y. Fu, K.Joo Kwak, Y. Fan, C. Kang, C. Yin, J. Rhee, P. Bertani, J. Otero, W. Lu, A.S. Lee, W. Jiang, L. Teng, B.Y.S. Kim and L.J. Lee, “Large-Scale Generation of Functional mRNA Containing Exosomes via Cellular Nanoporation”, Nature Biomedical Engineering, 4, 69-83 (2020).
Targeted Delivery by Lipoplex Nanoparticles- The high mortality rates for many cancers can be attributed to late diagnosis, high recurrence rates, metastasis, and limited effectiveness of current therapeutic modalities. Although the aforementioned issues have been studied extensively, there is no current technology platform that allows for simultaneous detection, therapy and prognosis determination. MicroRNA (miRNA) dysregulation has been implicated in HCC development. These miRNAs regulate a network of tumor promoting and tumor suppressor genes that are critical for tumor cell survival, epithelial-to-mesenchymal and mesenchymal-to-epithelial transition (EMT/MET), escaping the host immune system, resistance to therapy, and angiogenesis. Modulating tumor cells and its microenvironment with miRNA replacement (MRT) and anti-miRNA therapy (AMT) can potentially inhibit tumor growth and sensitize tumor cells to existing therapy. A critical barrier to the clinical development of MRT/AMT is that oligonucleotides are sensitive to nucleases, subject to renal and reticuloendothelial system (RES) clearance with minimum membrane permeability. Delivery systems based on targeted lipid nanoparticles can potentially address these problems. My team has made significant contribution in this area.
• B. Yu, Y. Mao, L. Bai, S. May, A. Ramanunni, Y. Jin, X. Mo, C. Carolyn, K.K. Chan, D. Jarjoura, G. Marcucci, R.J. Lee, J.C. Byrd, L.J. Lee and N. Muthusamy, “Liposomal Targeted Delivery Overcomes Off-target Immunostimulatory Effects of RNA Oligonucleotide”, Blood, 121, 136-147 (2013).
• X. Huang, S. Schwind, B. Yu, R. Santhanam, H. Wang, P. Hoellerbauer, A. Mims, R. Klisovic, A.R. Walker, K.K. Chan, W. Blum, D. Perrotti, J.C. Byrd, C.D. Bloomfield, M.A. Caligiuri, R.J. Lee, R. Garzon, N. Muthusamy, L.J. Lee and G. Marcucci, “Targeted Delivery of microRNA-29b by Transferrin Conjugated Anionic Lipopolyplex Nanoparticles: A Novel Therapeutic Strategy in Acute Myeloid Leukemia”, Clinical Cancer Research, 19(9), 2275-2292 (2013).
• Y. Wu, J. Ma, P Woods, N. Chesarino, L.J. Lee, S.P. Nana-Sinkam and I.C. Davis, DVM, “Selective Targeting of Alveolar Type II Respiratory Epithelial Cells by Anti-surfactant Protein-C Antibody-conjugated Lipoplexes”, J. Controlled Release, 203, 140-149 (2015).
• C-L Chiang, S. Goswami, F. Frissora, Z. Xie, P Yan, R. Bundschuh, L. Walker, X. Huang, R. Mani, X. Mo, S. Baskar, C. Rader, M. Phelps, G. Marcucci, J. Byrd, L.J. Lee, and N. Muthusamy, “ROR1-targeted Delivery of miR-29b Induces Cell Cycle Arrest and Therapeutic Benefit in vivo in CLL Mouse Model”, Blood, 134, 432-444 (2019).
Tethered Lipoplex Nanoparticle Biochips for Extracellular Vesicles Based Circulating RNA Biomarkers- Biomolecules such as miRNAs, lncRNAs, mRNAs and protein antigens can be useful as biomarkers for cancer and other diseases. Recent studies show that they are excreted by cells in the form of extracellular vesicles (EVs) including exosomes and can be detected in the blood. Our group has been actively engaged in establishing EVs as potential noninvasive biomarkers. EVs are known mobile elements that function as escape routes for proteins and RNAs from one cell (site of origin) to distant locations. Many studies have revealed that EVs cross talk and/or influence major disease-related pathways, such as hypoxia-driven EMT, angiogenesis, and metastasis, involving many cell types within the tumor microenvironment. However, existing methods based on next generation sequencing (NGS), hybridization microarrays, and qRT-PCR have limited sensitivity and require tedious and expensive sample preparation and detection procedures. They also need several hundred microliters of blood for EV analysis. This necessitates the sacrifice of animals in murine studies, and making it impractical for tumor monitoring in murine tumor model therapy trials. To address these issues, my lab has developed “tethered lipid nanoparticle (TLN)” technology that can be used for ultra-sensitive detection of miRNAs, lncRNA, mRNA and protein antigen targets in EVs. Analysis can be performed using only 20 µL of blood from a patient or mouse with minimal sample preparation requirement. This would enable “non-lethal” monitoring of tumor load in mice. We believe that serum/plasma EV miRNA/mRNA/membrane protein profiles can serve as detection, surveillance and prognostic biomarkers that can be used for early disease diagnosis and to monitor and predict disease response to therapy.
• Y. Wu, K.J. Kwak, K. Agarwal, A. Marras, C. Wang, Y. Mao, X. Huang, J. Ma, B. Yu, R.J. Lee, A. Vachani, G. Marcucci, J.C. Byrd, N. Muthusamy, K. Huang, C.E. Castro, M. Paulaitis, S.P. Nana-Sinkam and L.J. Lee, “Detection of Extracellular RNAs in Cancer and Viral Infection by Tethered Cationic Lipoplex Nanoparticles”, Analytical Chemistry, 85(23), 11265-11274 (2013).
• L.J. Lee, Z. Yang, M. Rahman, J. Ma, K.J. Kwak, J. McElory, K. Shilo, C. Goparaju, L. Yu, W. Rom, T-K Kim, X. Wu, Y. He, K. Wang, H.I. Pass and S.P. Nana-Sinkam, “Extracellular mRNA Detected by Tethered Lipoplex Nanoparticle Biochip for Biomarker Development in Lung Cancer”, American Journal of Respiratory and Critical Care Medicine, 193(12), 1431-1433 (2016).
• Hu, Y. Sheng, K.J. Kwak and L.J. Lee, “A Signal-amplifiable Biochip Quantifies Extracellular RNAs for Early Cancer Detection", Nature Communication, 8(1),1683 (2017).
• J. Hu, K.J. Kwak, Y. Sheng and L.J. Lee,” Overhang Molecular Beacons Encapsulated in Tethered Cationic Lipoplex Nanoparticles for Detection of Single-point Mutation in Extracellular Vesicle-associated RNAs”, Biomaterials, 183:20-29 (2018).
Cell-Based Drug Delivery- Cell-based therapeutic strategy has been proposed as a tissue engineering application for several decades. Primary or cell line with certain product secretion can be used as “seed” cell for therapeutic function. Alternative resource of cells is from gene recombination-mediated methods. As most tissue or cellular transplants, the cellular grafts are subject to immunorejection in the absence of chronic immunosuppression. For cell-based device applications in vivo, the device must provide proper cell immunoprotection with minimal inflammatory response. Furthermore, the device should possess controllable degradation characteristics such that the implant does not need to be removed after use via invasive second surgery. My lab has conducted considerable research in this area.
• X. Zhang, H. He and L.J. Lee, “A Biodegradable, Immunoprotective, Dual Nanoporous Capsule for Cell-Based Therapies”, Biomaterials, 29, 4253-4259 (2008).
• H. He, V. Grignol, V. Karpa, C. Yen, K. Laperle, X. Zhang, N.B. Jones, M.I. Liang, G.B. Lesinski, W. Ho, W.E. Carson, III and L.J. Lee, “Use of a Nanoporous Biodegradable Miniature Device to Regulate Cytokine Release for Cancer Treatment”, Journal of Controlled Release, 151, 239-245 (2011).
• F. Yang, X. Zhang, A. Maiseyeu, G. Michai, R. Yasmeen, D. DiSilvestro, K.S., Maurya, M. Periasamy, V. Bergdall, C. Sen, S. Roy, L. J. Lee, S. Rajagopalan, and O. Ziouzenkova, ”The Prolonged Survival of Fibroblasts with Forced Lipid Catabolism in Visceral Fat Following Encapsulation in Alginate-poly-L-lysine”, Biomaterials, 33(22), 5638-5649 (2012).
• H. He, E. Luedke, X. Zhang, B. Yu, A. Schmitt, B. McClarren, V. Grignol, W.E. Carson III and L.J. Lee, “A Nanoporous Cell-Therapy Device with Controllable Biodegradation for Long-Term Drug Release”, Journal of Controlled Release, 165(3), 226-233 (2013).
Polymer Based Micro/Nanofluidics Biochips- Micro/nanotechnology is initiated from the electronics industry. It has been extended to micro-electro-mechanic system (MEMS) and nano-electro-mechanic system (NEMS) for producing miniature devices based on silicon and semi-conductor materials. However, the use of these hard materials alone is inappropriate for many biomedical devices. Soft polymeric materials possess many attractive properties such as high toughness and recyclability. Some possess excellent biocompatibility, are biodegradable, and can provide various biofunctionalities. Proper combinations of micro/nanoelectronics, polymers, and biomolecules can lead to new and affordable medical devices. My team has established a series of
non-cleanroom and cleanroom manufacturing techniques using biocompatible polymers, biomolecules, and nanoparticles as building blocks as well as micro/nanofluidics as a mechanism to design, synthesize, and fabricate biomedical and therapeutic devices. In addition to drug/gene delivery devices and cell-based constructs described earlier, we have also developed various biosensors/chips using advanced micro/nanofluidics concepts for medical diagnostics.
• J. Guan and L.J. Lee, “Generating Highly Ordered and Stretched DNA Arrays”, Proceedings of National Academy of Science, 102(51), 18321-18325 (2005).
• S. Wang and L.J. Lee, “Dynamic Assembly by Electrokinetic Microfluidics”, Journal of American Chemical Society, 129(2), 254-255 (2007).
• Y. Yang, D. Liu, L.J. Lee, and D.L. Tomasko, “Low Temperature Fusion of Polymeric Nanostructures Using Carbon Dioxide”, Advanced Materials, 19(2), 251-254 (2007).
• P. Boukany, O. Hemminger, S-Q Wang and L.J. Lee, “Molecular Imaging of Slip in Entangled DNA Solution”, Physical Review Letters, 105(2), 027802 (2010)
BREAKTHROUGH GENE THERAPY: INJECTABLE, PROGRAMMABLE NANOCARRIERS FOR CUSTOM MEDICINE DEVELOPMENT OF A GENERAL PLATFORM TO PURIFY APO-PROTEINS
Innovation: Jim Lee’s gene-therapy method transforms human cells into mass producers of tiny nano-sized particles containing genetic material with the potential to reverse disease processes, kill cancer cells or regenerate organs, or silence or activate specific genes.
Impact: Potential application in therapeutics that surmounts bio-barriers: Deliverable to any target within the body, including safely accessing the brain, without provoking an immune response.
UNPRECEDENTED EARLY-STAGE CANCER DETECTION: NOVEL EV DIAGNOSTICS IN LIQUID BIOPSY
Innovation: Lee, in conjunction with Eduardo Reátegui, is developing a minimally invasive liquid biopsy that could transform cancer diagnosis and treatment. Impact: Provides an accurate and efficient analysis of molecular content within individual extracellular vesicles (EVs) from bodily fluids, creating a faster and pain-free bop[sy process.
Shared Facility Highlights
- Dynamic light scattering goniometry
- Total internal reflection fluorescence (TIRF) microscopy
During his long and highly productive career, Professor Lee raised more than $120 million in research grants from federal and local government agencies as well as industry.
A Mother Nature-induced therapeutic nanoparticle developed by Lee group
Professor Lee has an extensive record of publishing, including
- More than 400 papers with more than 15,000 citations
- Publications with IF>10:
- 3 in Nature Nanotechnology,
- 1 in Nature Biomedical Engineering,
- 1 in Nature Communication,
- 2 in Cancer Cell,
- 6 in Advanced Materials,
- 5 in Blood,
- 1 in American Journal of Respiratory and Critical Care Medicine,
- 1 in Proceedings of National Academy of Science,
- 2 in Journal of American Chemical Society,
- 8 in Biomaterials,
- 3 in ACS Nano,
- 1 in Nano Letters,
- 1 in Clinical Cancer Research,
- 4 in Small,
- 2 in Physical Review Letters
94. C. Yen, H. He, Z. Fei, X. Zhang, L.J. Lee, and W.S. Ho, "Surface Modification of Nanoporous Poly( -caprolactone) Membrane with Poly(ethylene glycol) to Prevent Biofouling: Part II. Effects of Graft Density and Chain Length", International Journal of Polymeric Materials, 59(11), 943-957 (2010).
PATENTS AND PATENT APPLICATIONS (17 licensed to industry)
1. C.W. Macosko and L.J. Lee, "Reaction Injection Molding Machine," U.S. Patent 4,189,070, February 19 (1980).
2. L.J. Lee, J.F. Stevenson and R.M. Griffith, "Method of Producing an Extrudate Having Controlled Shape and Size," U.S. Patent 4,425,289, January 19 (1984).
3. L.J. Lee, R. Saito and Y.Y. Chiu, "Modification of Unsaturated Polyester Resins for Viscosity Control," U.S. Patent 5,561,192, October 1 (1996).
4. W.D. White, M.A. Sagai, L.J. Lee, and K. Han, "System and Method for Modeling Plastic Molding and Molding Parts Incorporating the SAME," U.S. Patent 5,581,468 (1996).
5. L. J. Lee, K.W. Koelling, D.L. Tomasko, X. Han and C. Zeng, “Polymer Nanocomposite Foams”, U.S. Patent 6,759,446, July 6 (2004).
6. R.R. Loh, M.E. Polasky, Y. Delaviz, L.J. Lee, X. Cao, J. Shen and B. Patel, “Polystyrene Foam Containing a Modifier-Free Nanoclay and Having Improved Fire Protection Performance”, U.S. Patent 11/221,522 (2005).
7. L. J. Lee, K.W. Koelling, D.L. Tomasko, X. Han and C. Zeng, “Polymer Nanocomposite Foams”, U.S. Patent 7,026,365, April 11 (2006).
8. L.J Lee and S. Lai, “Gas-Assisted Resin Injection Technique for Bonding and Surface Modification of Microfluidic Devices”, U.S. Patent 7,122,093, October 17 (2006).
9. L.J. Lee and C. Zeng, “Clay Nanocomposites Prepared by In-Situ Polymerization”, U.S. Patent 07,129,287B1, October 31 (2006).
10. J. Guan, L.J. Lee and D. Hansford, “Fabrication of Polymeric Micro-Devices for Drug Delivery by Surface Patterning and Self-Folding”, U.S. Patent 7,364,675, April 29 (2008).
11. L.J. Lee, D.L. Tomasko, Y. Yang and C. Zeng, “Carbon Dioxide Assisted Processing and Bonding of Polymer and Polymer Composites”, U.S. Patent 7,501,039, March 10 (2009).
12. R.R. Loh, M.E. Polasky, J.P. Rynd, L.J. Lee, X. Han and K.W. Koelling, “Polymer Foams Containing Multi-functional Layered Nano-graphite”, U.S. Patent 7,605,188, October 20 (2009).
13. A. Epstein, L.J. Lee, C. Lu and N-R Chiou, “Fabrication Methods and Applications of Alligned and Oriented Nanofibers of Polyaniline and Their Derivatives”, U.S. Patent 8,038,907, October 18 (2011).
14. L.J. Lee, G. Zhou and X. Cao, “A Method of Preparing a Composite with Disperse Long Fibers and Nanoparticles”, U.S. Patent 8,143,337, March 27 (2012).
15. A. Epstein, L.J. Lee and N-R Chiou, “Aligned Nanostructured Polymers”, U.S. Patent 8,293,140, October 23 (2012).
16. L.J. Lee, J. Yang, S-K Yeh and N-R Chiou, “Suspension Polymerization and Foaming of Water Containing Activated Carbon-Nano/Microparticulate Polymer Composites”, U.S. Patent 8507568B2, August 13 (2013).
17. L.J. Lee, D. Guerra, S Movva and Y.G. Min, “Layer-by-Layer Fabrication of Sprayed Nanopaper”, U.S. Patent 8,574,677, November 5 (2013).
18. L.J. Lee, K.J. Kwak, B. Yu and Y. Wu, “Tethered Lipoplex Nanoparticle Microarray Containing Molecular Probes for Cell, Microvesicles, Exosomes and Virus Capture and Detection”, China Patent CN103197066B on December 23 (2015).
19. L.J. Lee, P. Boukany, J. Guan and N. Chiou, “Dose and Location Controlled Drug /Gene/Particle Delivery to Individual Cells by Nanoelectroporation”, U.S. Patent 9,816,086, November 14 (2017).
20. L.J. Lee, J. Yu and Y-C Yen, “Graphene-like Nanosheet Structure Network on a Substrate and the Method for Forming the Same”, China Patent CN105274489B on June 19 (2018).
21. L. J. Lee, K. J. Kwak, B. Yu, Y. Wu and A. Lee, “Tethered Lipoplex Nanoparticle Biochips and Methods of Use”, U.S. Patent 10705085B2, July 7 (2020).
22. J. Byrd, N. Muthusamy, R. Lee, L.J. Lee, R. Lapalombella, B. Yu, “Methods and Compositions for Delivering Therapeutic Agents in the Treatment of B-Cell Disorders”, U.S. Patent Application No. 13/056,009 filed on January 26 (2011).
23. N-R Chiou, L.J. Lee, J. Yang and S-K Yeh, “Surfactant-Free Synthesis and Foaming of Liquid Blowing Agent-Containing Activated Carbon Nano/Microparticulate Polymer Composites”, U.S. Patent Application No. 12/995,042 filed on June 19 (2011).
24. W. Huang, Y Min and L.J. Lee, “Functionalization of Carbon and Non-carbon Particles and Their Applications”, U.S. Provisional Patent Application No. 61/629,939 on December 1 (2011).
25. L.J. Lee, W. Huang and J. Yu, “Covalently-Bonded Graphene Coating and Its Applications Therefore”, U.S. Patent Application PCT No. 14/409,815 on December 19 (2014).
26. L.J. Lee, J. Yu and Y-C Yen, “ Graphene-like Nanosheet Structure Network on a Substrate and the Method for Forming the Same”, U.S. Patent Application No. 14/335,072 on July 18 (2014).
27. L.J. Lee, K.J. Kwak, A. Lee, “Immuno Lipoplex Nanoparticle Array Biochip Containing Molecular Probes for Capture and Characterization of Extracellular Vesicles”, U.S. Patent Application No. 14/992,169 on January 11 (2016).
28. L.J. Lee, Y-C Yen, T. Lee, D. Chiu and C.S. Lin, “Semi-crystalline Polymer Nanocomposite and Foam Structure and Method for Making the Same”, TW patent Application No. 105121340 on July 6; U.S. Patent Application No.15/220,634 on July 27; CN patent Application No. 201610598896.70 on July 27 (2016).
29. L.J. Lee, K.J. Kwak, B. Yu, Y. Wu and A. Lee, “Tethered Lipoplex Nanoparticle Biochips and Methods of Use”, U.S. Patent Application PCT No. 15/279,545 on September 29 (2016).
30. L.J. Lee, J. Hu and K.J. Kwak, “Highly Stable and Specific Molecular Beacons Encapsulated in Cationic Lipoplex Nanoparticles and Application thereof”, U.S. Patent Application No. 15/880,309 on January 25 (2018).
31. C. Sen, L.J. Lee, D. Gallego-Perez, D. Pal and S. Ghatak, “Compositions and Methods for Reprogramming Somatic Cells into Induced Endothelial Cells”, U.S. Patent Application Patent Application No. 62/530,132 on July 8 (2017).
32. L.J. Lee, Z. Yang and J. Shi, "Method for Producing Therapeutic Exosomes from Nanoelectroporation and Other Non-endocytic Cell Transfection", U.S. Provisional Patent Application No. 62/541,157 on August 4 (2017).
33. L.J. Lee and J. Shi, "A Microfluidic-Nanoelectroporation Based Cell Transfection Method", U.S. Provisional Patent Application No. 62/631,251 on February 15 (2018).
34. L.J. Lee, Z. Yang and J. Shi, “Method for Producing Therapeutic Exosomes from Nanoelectroporation and Other Non-endocytic Cell Transfection”, U.S. Patent Application Patent Application No. 16/986,954 on August 6 (2020).
35. L.J. Lee, C-L Chiang and Y. Ma, “Adapter Polypeptides and Methods of Using the Same”, U.S. Provisional Patent Application No. 63/061,749 on August 5 (2020).
36. L.J. Lee, Z. Yang and J. Shi, “Therapeutic Extracellular Vesicles”, U.S. Patent Application Patent Application No. 16/986,954 on August 6 (2020).
37. L.J. Lee and K.J. Kwak, “Extracellular Vesicle Methods”, U.S. Provisional Patent Application No. 63/133,735 on January 4 (2021).