Wood Laboratory for Applied Protein Engineering
Wood Laboratory for Applied Protein Engineering
Biotechnology development through protein engineering
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About Us
Led by Professor David W. Wood, our work seeks to develop highly useful biotechnologies through engineering proteins for specific applications. So far, these applications include new ways to purify recombinant proteins, bacterial biosensors that incorporate human drug targets, and new capabilities in drug discovery and drug delivery.
- Learn more about the Applied Protein Engineering Group in the video at the left.
- To view in full-screen mode, click on the Play button and then on the square in the bottom right of the video.
- Professor Wood is currently accepting students.
Faculty Bio
In Brief
Since receiving his PhD from Rensselaer Polytechnic Institute (RPI) in 2001, Professor Wood has developed groundbreaking new technologies in downstream bioprocessing, with a focus on self-cleaving affinity tag methods. His work has led to several issued patents in this area, and he is now an active consultant in the area of biosimilars development and intellectual property, as well as a co-founder of Protein Capture Science, LLC, a startup company commercializing his self-cleaving affinity tag technology.
Key Distinctions
- Seven patents
- Developed self-cleaving affinity tag technology for the purification of recombinant proteins
- Founder, Protein Capture Science, LLC. Wood's patents and main sales product of his startup company comprises a new platform technology for purifying recombinant proteins, which have the potential to accelerate research and simplify the manufacture of therapeutic proteins. The result would be faster patient care and new and cheaper biopharmaceuticals entering the market.
- National Science Foundation CAREER Award, 2003
Expertise
Novel and/or non-chromatographic recombinant protein bioseparations; pharmaceutical bioprocess development; incorporation of functional human drug targets into bacterial reporter proteins; design of controllable, self-activating proteins for medical and research applications; detection and identification of environmental endocrine disrupting compounds.
Self-Cleaving Affinity Tag Technology
Professor Wood's research focuses on biotechnology development through protein engineering. He is known for groundbreaking research in self-cleaving affinity tag technology for the purification of recombinant proteins. Current applications include new ways to purify recombinant proteins, bacterial biosensors that incorporate human drug targets, and new capabilities in drug discovery and drug delivery.
Since the mid-1980s, the use of affinity tag technology has been a ubiquitous method for laboratories to purify virtually any arbitrary recombinant protein through a single general and simple method. While dozens of different tags, kits and accessories are now commercially available, this method has not been adopted for large-scale bioprocessing, largely due to the expense of removing the affinity tag after the target protein is purified.
The Wood Lab is currently developing new protein elements that effectively make the affinity tags self-cleaving after purification, therefore enabling many new tag-based protein purification methods that address the biotech industry's needs for scale, simplicity and cost control.
Background
Professor Wood's interest in protein engineering started when he was an undergraduate at Caltech, where he received a dual degree in Chemical Engineering and Molecular Biology in 1990.
During this time, he did undergraduate research in the laboratories of Frances Arnold in Chemical Engineering, and William A. Goddard III, in Chemistry and Applied Physics. Both of these professors are well-known in protein engineering and protein structure prediction.
Wood then worked one year at Kelco in bioprocess development for commodity-scale mucopolysaccharide products, and two years at Amgen Inc. working with the Neupogen® manufacturing process.
After these industrial experiences, he returned to graduate school at RPI in 1993, where he was co-advised by Georges Belfort (at RPI) and Marlene Belfort (at the NY State Dept of Health, Division of Genetic Disorders).
During his graduate work, he combined rational protein engineering with evolutionary approaches to develop a novel self-cleaving intein for applications in protein purification. This intein provides a means to make any protein purification tag self-cleaving and has now been patented (US patent # 6,933,362) and incorporated into several new technologies.
Wood completed his PhD in 2001 and joined the faculty at Princeton University in the Department of Chemical Engineering. At Princeton, he continued working in protein engineering to develop new biotechnologies at the molecular level. This work now continues at The Ohio State University, where an important advance has been the development of several new self-cleaving tag modules that allow proteins and protein complexes to be purified in a variety of formats, and a new effort in split-intein methods.
As of April 2021, Wood's work in engineered proteins for biosensing and bioseparations has led to 60+ peer-reviewed journal papers, 11 book chapters, and one co-edited volume, in addition to over 40 invited conference presentations and seminars at prestigious institutions.
Strains developed in Wood's laboratory have also been requested by over 180 researchers worldwide, primarily for applications in bioseparations.
Research
The Applied Protein Engineering Group's research focus is on biotechnology development through protein engineering. Professor Wood is known for groundbreaking research in self-cleaving affinity tag technology for the purification of recombinant proteins. Current applications include new ways to purify recombinant proteins, bacterial biosensors that incorporate human drug targets, and new capabilities in drug discovery and drug delivery.
Since the mid-1980s, the use of affinity tag technology has been a ubiquitous method for laboratories to purify virtually any arbitrary recombinant protein through a single general and simple method. While dozens of different tags, kits and accessories are now commercially available, this method has not been adopted for large-scale bioprocessing, largely due to the expense of removing the affinity tag after the target protein is purified.
The group is currently developing new protein elements that effectively make the affinity tags self-cleaving after purification, therefore enabling many new tag-based protein purification methods that address the biotech industry's needs for scale, simplicity and cost control.
Prospective students, postdoctoral researchers, and visiting scholars should reach out to Dr. Wood directly for more information on incoming projects in any of these areas.
Bioseparations
Our lab group uses both mammalian and bacterial systems to express proteins of interest (E. coli, bacillus; HEK293, Expi293, ExpiCHO). Many of these systems are used by biopharmaceutical companies to produce therapeutic proteins, giving our students the opportunity to train in techniques desirable to future employers. Bacterial expressions are performed in our BSL1 laboratory space using shake flasks with capacity up to 10L/day. Alternatively, mammalian expressions are done in a BSL2 certified room within the lab. Some current projects using bacterial systems are expressing recombinant hemoglobin, designing a reusable protease, and various protein engineering projects. Mammalian proteins that we are currently working with include monoclonal antibodies, bispecific antibodies, scFvs, SARS-CoV2 RBD, aminoacyl-tRNA synthetase (EPRS), and others.
We have developed new protein elements that effectively make the affinity tags self-removing after purification, and therefore enable many new tag-based protein purification methods. These methods retain the simplicity and generality of conventional affinity tag technology and can potentially make affinity tags a ubiquitous and critical platform technology in industrial protein bioseparations. We have commercialized our most promising self-removing affinity tag (iCapTag TM) based on the naturally occurring Npu DnaE split intein and demonstrated its use for purifying proteins from multiple host cells. Due to its small size and minimal impact on protein solubility, iCapTag TM can serve as a potential platform technology for the purification of diverse classes of therapeutic proteins. Further, using protein engineering principles, we have been able to make iCapTag TM pH sensitive which allows us to suppress the cleaving reaction at high pH (>8.0) enabling the use of high salt washes to remove impurities. Once the pH is shifted to mildly acidic (~ 6.0), the cleaving reaction is accelerated, and we obtain tagless product. We have also shown that iCapTag TM can be used in tandem with other affinity tags to perform dual affinity column processes to achieve high clearance of process and product related impurities.
Current projects focus on the development of newer self-removing affinity tags with faster cleaving kinetics, higher binding capacities and better controllability. A few papers summarizing our work on this area over the last two decades have been shown below.
- Wood, D., Derbyshire, V., Wu, W., Chartrain, M., Belfort, M. and Belfort, G., “Optimized Single-Step Affinity Purification with a Self-Cleaving Intein Applied to Human Acidic Fibroblast Growth Factor,” Biotechnology Progress, Vol. 16, pp. 1055-1063, (2000).
- Wu, W., Wood, D., Belfort, G., Derbyshire, V. & Belfort, M., “Intein-Mediated Purification of Cytotoxic Endonuclease I-TevI by Insertional Inactivation and pH-Controllable Splicing,” Nucleic Acids Research, Vol. 30, pp. 4864-71, (2002).
- Coolbaugh, M. J., Shakalli Tang, M. J., Wood, D. W., “High-throughput purification of recombinant proteins using self-cleaving intein tags,” Analytical Biochemistry, Vol. 516, pp. 65-74, (2017).
- Fan, Y.; Miozzi, J.M., Stimple, S.D., Han, T.-C., Wood, D.W., “Column-Free Purification Methods for Recombinant Proteins Using Self-Cleaving Aggregating Tags,” Polymers, Vol. 10, pp. 468 (2018).
Proteins engineering and bioprocess development requires ample biochemical characterization to further comprehend molecular systems. By using different biophysical techniques such as differential scanning fluorimetry (DSF), isothermal titration calorimetry (ITC), circular dichroism (CD), surface plasmon resonance (SPR), and other techniques, more data can be acquired to build a molecular model. DSF is a high throughput method that assess the protein thermal denaturation and how modifications in the protein could that impact the stability of the final purified and formulated product. CD provides information about the protein secondary structure elements, while ITC helps to understand protein-ligand and protein-protein interactions by measuring heat changes while titrating a binding molecule. This method can also generate bulk thermodynamic parameters such as free energy, entropy, stoichiometric factor, and binding affinity of the protein model. SPR can be used to determine the association and dissociation kinetic rates in real-time. These techniques can inform about protein stability, folding, binding, and other factor to further validate the student’s experimental designs and translate them into applicable biotechnologies, and manufacturing platforms for scale up processes in industry.
Biosensing and Drug Discovery
With the newly developed self-cleaving intein tag, the laboratory's research has been primarily focused on applying this technology to downstream processing, particularly in the biopharmaceutical industry. Through his research, Dr. Wood has gained substantial awareness and expertise in the potential applications of tag technologies in biopharmaceutical development and manufacturing. Our group has had extensive contact, discussions, and sponsored research projects with major companies including Merck (USA), Miilipore (Germany), Evox (UK). Aside from the intein technology, our lab supports research in major chromatographic methods (affinity, ion-exchange, other) in both bulk/gravity and AKTA FPLC modalities. An example of recent work in this area includes the optimization of a column-free bulk binding protocol which has been accepted for publication in Current Protocols in Protein Science (listed below with other recent publications in this area).
- Cooper, M. A.,* Taris, J. E.,* Shi, C. and Wood, D. W., “A convenient self-cleaving affinity tag method for the purification of tagless target proteins (Invited Methods Paper),” Current Protocols in Protein Science, Vol. 91, pp. 5.29.1-5.29.23, (2018).
- Gierach, I., and Wood, D. W., “Self-Cleaving Tags Based on Split Inteins: Increased Reliability Enabling Higher Throughput Applications,” American Pharmaceutical Review, Vol. 22 (1), pp. 66-69, (2019).
- Prabhala, S.V., Mayone, S.A., Moody, N.M., Kanu, C. B. & Wood, D. W. (2023). Convenient split-intein tag methods for the single step purification of tagless target proteins. Current Protocols in Protein Science, 91, 5.29.1–5.29.23. doi: 10.1002/cpps.46
We have developed expertise in applied protein engineering for specific applications. In most cases, our approach involved the recombination of protein subdomains to generate useful fusions with beneficial properties. In the past, we have used this strategy to generate allosteric biosensors for endocrine disrupting compounds, complex tags for various strategies in protein detection and purification, and more recently in the development of our split intein affinity tag platform. More exotic applications have involved the engineering of new and improved activities into existing proteins and enzymes, where a recent project centers on the development of hydrolytic activity in a binding protein through the introduction of catalytic metal centers. These methods combine both rational protein engineering based on knowledge of the protein structure and active site, along with some evolutionary methods to improve desired catalytic activity. A key aspect of all of this work is that we have the goal of generating proteins with specific needed applications in biotechnology and medicine.
Enzyme-polymer nanoparticle formulations are advanced biomanufacturing technology offering a versatile platform with a wide range of industrial applications. As a multidisciplinary approach, this research connects protein engineering with polymer chemistry and molecular biology to develop a model series of polymer-based enzymatic nanomaterials. We have designed self-assembling amphiphilic diblock copolymers as versatile, non-toxic, cost-efficient nanoplatforms for enzyme immobilization. This nanoenzyme formulation involves bioconjugation (physical/covalent) of recombinant enzymes with biocompatible and biodegradable polymer nanoparticles derived from agro-industrial wastes. These polymer-based nanoplatforms have been highly effective in enhancing enzyme properties (i.e., stability, recovery, and specificity) with cost-competitive enzymatic nanomaterials applications, especially in the food, agriculture, and energy industries.
Link List
Tiles
Additional Lab Members and Alumni
Current
- Farah Deeba Ph.D., Postdoctoral Scholar, enzyme-based pesticides and nanoparticles, 2018-current
Former
- Tarek Mazeed, Visiting Scholar, fermentation development
- Richard Lease, Ph.D., Research Scientist, metabolic engineering for biofuels production
- Samar Al-Sharawi, Visiting Scholar, thermophilic enzyme isolation
- Xia Hai-Feng, Visiting Scholar, intein Purification methods
- Changhua (Steven) Shi, Co-advised Ph.D. and Postdoctoral Scholar, Split inteins. Last known position: Senior Scientist, WuXi Biologics, Shanghai, China
- Junfen Wan, Ph.D., Visiting Scholar, Defence Advanced Research Projects Agency (DARPA) project on protein manufacturing. Last known position: Assoc. Prof. & Ph.D., School of Biotechnology, East China University of Science and Technology, Shanghai, China
- Jingjing Li, Postdoctoral Scholar, development of a bacterial screen for potential autism-related compounds.” Last known position: Postdoctoral Research Associate, UNC, Chapel Hill, NC
- Iraj Ghazi, Postdoctoral Scholar, Research Scientist, development of viable carbonic anhydrase enzyme process for carbon sequestration and DARPA "BioMOD." Last known position: Bioprocess Development Specialist II, Shire Pharmaceuticals, Boston, MA.
- Dr. Sai Vivek Prabala, "“Advances in affinity-based methods for downstream process development of monoclonal antibody and recombinant protein therapeutics," Ph.D. granted December 2023. Last known position: Senior Scientist, Merck
- Dr. Hongyu Yuan, "“Remarkable Advances in Developing Contiguous Intein as Multi-functional Tools in Application of Downstream Purification and Drug Delivery,” Ph.D. granted July 2023. Last known position: seeking employment
- Dr. Steven Cummings, "“In vitro Reconstitution of Tubulin Polyglycylation," Ph.D. granted May 2023. Last known position: Patent technical Specialist, Medler Ferro Woodhouse & Mills PLLC
- Dr. Brian Marshall, "“Development and Applications of a Next-Generation Split-Intein Based Affinity Chromatography Ligand and a Characterization of the Effects of the N-exteins on the Cleaving Kinetics of a Contiguous Intein," Ph.D. granted April 2022. Last known position: Virology and purification development, Eli Lilly
- Dr. Joseph Taris, "Development of a Novel Intein-Mediated Affinity Capture Platform for Production of Recombinant Proteins and Biopharmaceuticals," Ph.D. granted September 2021. Last known position: unknown
- Dr. Jackelyn (Miozzi) Galiardi, "Split Intein Applications for Downstream Purification and Protein Conjugation," Ph.D. granted April 2021. Last known position: Senior scientist, Pfizer
- Dr. Kevin McGarry (through OSBP), “Reengineering Butyrylcholinesterasefor the Catalytic Degradation of Organophosphorus Compounds”, Ph.D. granted May 2019. Last known position: cGMP Quality Control Supervisor - Clinical Manufacturing, Nationwide Children's Hospital.
- Dr. Yamin Fan, “Split Inteins As Versatile Tools in Applications of Downstream Purification and Bioconjugation”, Ph.D. granted May 2019. Last known position: Sr. Engineer I, Biogen.
- Dr. Samuel Stimple, “Recent Advances in Developing Molecular Biotechnology Tools for Metabolic Engineering and Recombinant Protein Purification”, Ph.D. granted August 2018. Last known position: Scientist, Antibody Research, Sanofi.
- Dr. Merideth Cooper, “Creating an Efficient Biopharmaceutical Factory: Protein Expression and Purification Using a Self-Cleaving Split Intein”, Ph.D. granted May 2018.Last known position: Advanced R&D Engineer at Owens Corning.
- Dr. Ashwin Lahiry, “Developing Molecular Tools for Applications in Metabolic Engineering and Protein Purification.” Ph.D. granted August 2017. Last known position: Research Scientist, Merck, Inc.
Thesis
- Angela Chen (NSF GFRP), University of Texas PhD program. Last known position: Postdoctoral scholar at UC-Riverside.
- Dr. Hannah Zierden (NSF GFRP), Last known position: Assistant Professor, Chemical and Biomolecular Engineering, University of Maryland
- Dr. Robert Law, Sr.. Last known position: Senior Immunoassay Scientist, AOA Dx
- Robert Pittman, Ohio State University Masters Program
- Brian Novi, Philadelphia College of Osteopathic Medicine
- Robert Wensing, University of Illinois, Urbana Champaign PhD program
Non-Thesis
- Jordan Loeffler (Co-op, downstream process development, Merck)
- Reva Sharma (Intern, analytical sciences, Dow)
- Emily Wiegand (Analyst at J.P. Morgan)
- Haley Scott (Intern, Forge Biologics)
- Mason Pierce (Co-op, preclinical manufacturing and process development analytics, Regeneron Pharmaceuticals)
- Dominic Cirillo (Intern, computational methods, Moderna)
- Mira Faizul (Intern, Sarepta Therapeutics)
- Caleb Russel (Intern, Forge Biologics, Co-op, materials science, Battelle)
- Natalie Hoffman (Intern, biologics analytical R&D, Merck, Intern, materials science, Battelle)
- Sathvik Kethireddy (Johns Hopkins University Masters program)
- Emery Monnig (Mt. Saini M.D. program)
- Sophia Krebs (Scientist, Forge Biologics)
- Yara Mohamad (Manufacturing Associate at ThermoFisher)
- Olivia Krebs (NSF GFRP), Doctoral student, Case Western Reserve University BME Ph.D. program
- Matthew Haynam (Bioprocess Engineer at AveXis, a Novartis Company)
- Maria DeBastiani (Process Development Associate, Regeneron Pharmaceuticals)
- Maria Znidarsic (Engineer, Biogen Corporation)
- Jonas Immel-Brown (Research Scientist, Biogen Corporation)
- Hopen Yang (University of Delaware Ph.D. program)
- Jacob Martin (Northwestern University Ph.D. program)
- Jesse Westfall (Clemson University Ph.D. program)
- Connor Weigand (University of Pittsburgh Ph.D. program)
- Mitch Raith (University of Tennessee at Knoxville Ph.D. program)
- Tiana Warren (NSF GFRP) (Johns Hopkins PhD program; Scientist II at BD Corp.)
- Jesse Leissa (University of Maryland M.S. program; R&D Associate II at AstraZeneca)
- Jonathan Strutz (Northwestern University PhD program)
- Alexis Brannan (B.S., Purdue University; Advanced Product Assurance Engineer at 3M)
- Lily Glick (Staff Engineer at Kenexis Consulting Corporation; J.D. Candidate, OSU Law School David Cain, Associate engineer at Genentech)
- Akshit Gupta (Analyst at Clinton Health Access Initiative, New Delhi Area, India)
- Jason Porter (Quality Assurance Manager at Starr Hill Brewery, Charlottesville, Virginia)
- Brian Saunders (Territory Manager II - OH/MI Packaging - at H.B. Fuller)
- David Benco (Customer Support Engineer, OSIsoft)
- Derek Reichel (University of Kentucky PhD program)
- Gina Pietro (Field Engineer at Baker Hughes Geoffrey Kleimeyer, Dow Chemical)
- Theodore Rader (M.D., University of Toledo; Resident Physician at VCU Health)
- Alnuel Barnum
- Alex Miller
- Kyle Schneider
- Madison (Matti) Srock
- William (Will) Dixon
- Will Caines
- Martin (Marty) Evard
- Max Kleinmann
- Joseph Orlando
- Anna Asim
- Noah Breitenbecher
- Steven Youn
- Christy Caporale
- Aniliese Deal
- Emily Doleh
- Elizabeth Judy
- Jasur Khan
- Manasa Korrapati
- Laura Kryah
- Natalya Lavrenchuk
- Orion Wang
- Connor Weyrick
- Brandon Clem
- Don Matthew
- Elias Linhardt
- Jim Bradcovich
- Joel Silleck
- Joel Francis
- Isaac Delev
- Yujing Zhai, “Evaluation of a Zinc Binding-domain for Control of Intein Cleaving.” MS granted November 2016. Last known position: Research Scientist, Genomics Institute of the Novartis Research Foundation, San Diego, CA
Facilities and Equipment
Lab facilities are located in the CBEC building, which was constructed in 2015 with state-of-the-art features.
Wood Lab has a wide variety of equipment facilitating biopharmaceutical research. Interested students, visiting scholars and collaborators should contact Professor David Wood at wood.750@osu.edu and visit the Chemical and Biomolecular Engineering homepage.
Industrial and private clients are likewise encouraged to contact Professor David Wood to see how our lab can develop solutions to address your bioprocessing needs.
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Our Lab in Ohio State News
News from the Wood Laboratory
News from the Wood Laboratory
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External Commentaries About Our Work
Genetic Engineering & Biotechnology News (GEN) | "Microbial Culture Systems for Bioprocessing," May 2019.
- Wood was a featured expert discussing the challenges and choices in microbial production, including type of organism, optimization of medium, and upscaling of production systems.
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Research Media Europe | International Innovation: Bacterial Biosensors: A Conversation with Dr. David Wood
- How biosensing techniques can help in drug discovery by identifying compounds that mimic or inhibit hormones that instigate health disorders
- Understanding Autism Spectrum Disorder
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Chemistry World | Grabbing Proteins by the ELPs
- A breakthrough method for protein purification based on self-cleaving polypeptide tags promises to be simpler and cheaper than affinity chromatography, the current method for purifying recombinant proteins.
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Protein Science | Cold Spring Harbor Laboratory Press: Proteins from PhB granules
- George Georgiou and Ki Jun Jeong, University of Texas, Institute for Cellular and Molecular Biology, Department of Biomedical Engineering, comment on Wood's research.
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Nature Biotechnology | Fine-Tuning an Engineered Intein
- Description of an ‘elegant’ mutational strategy to engineer an intein with improved features to serve as a tool for protein purification.
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Science Online | Melding Talents with a Career in Bioprocessing
- Discussion of today’s bioprocessing spectrum by Dr. David Wood, and training and career opportunities in the field.
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Biotechnology and Bioengineering Spotlight: | Streamlining the Process from Gene to Pure Protein
- Two new methods that simplify the goal of cloning and expressing new target genes and purify their encoded proteins.
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The Scientist | Pure Protein Sans Columns
- How intein-based protein-purification avoids column chromatography.
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Active Funding
National Institutes of Health
- "Engineering a Novel Biomaterial for Oxygen Transport Applications." Project seeks to develop highly structured hemoglobin-based oxygen carriers for basic research in artificial blood design. Wood, PI; with Andre Palmer. Amount: $2,716,911 (2020-2024)
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"Engineering human butyrylchlinesterase for catalytic hydrolysis of organophosphorus nerve toxicants." This proposal seeks to develop a safe treatment for exposures to chemical warfare nerve agents and specific pesticides. Wood; PI; with Hannah Shafaat. Ammount: $203,908 (2023-2025)
Industry Sponsorship
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"Merck intein cleaving work." This proposal is to evaluate our self-removing tag technology for the development of a novel vaccine being developed at Merck. Wood; PI. Amount: $150,000 (2023-2024)
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"Evox intein tuning work." This proposal is a research contract to develop highly controlled inteins for use in exosome-based drug delivery tools with Evox Therapeutics. Wood; PI. Amount: $150,00 (2021-2023)
Ohio State Internal Awards
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"Polymer-based enzymatic nanomaterials." This proposal seeks to develop renewable, biodegradable and edible nanoparticles for enzyme immobilization and stabilization. Wood; PI with Davita Watkins. Amount: $70,000 (2023-2024)
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"Enzymatic Nanofungicide as an Innovative Food Packaging Biomaterial." This award is to support utilization of the IMR core facilities for characterizing our nanoparticles. Wood; PI with Davita Watkins. Amount: $2,500 (2024)
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"Stable, Sustainable, and Green Bioengineering and Biomanufacturing of Nanocatalysts." This award is to develop our nanoparticles and determine potential manufacturing processes for large-scale implementation under additional funding. Wood; PI with three co-PIs. Amount: $100,000 (2024-2025)
Links and Downloads
Chemical Engineering
- American Institute of Chemical Engineers (AIChE)
- Computing and Systems Technology (CAST) division of AIChE
Bioengineering Societies
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American Chemical Society - Division of Biochemical Technology
Bioprocess Engineering and Biopharmaceuticals
- Cambridge Innovation Institute - Chi Pep Talk
- Bioprocess International
- BioPharm International