John Slater, Associate Professor, University of Delaware, United States of America -- Conference Chairperson

Chairperson's Introduction and Welcome and Introduction to the Scope and Topics Addressed

**Introduction to Biofabrication and Manufacturing**

​

Ali Khademhosseini, CEO, Terasaki Institute for Biomedical Innovation, United States of America

Engineering in Precision Medicine

Engineered materials that integrate advances in polymer chemistry, nanotechnology, and biological sciences have the potential to create powerful medical therapies. Dr. Khademhosseini is interested in developing ‘personalized’ solutions that utilize micro- and nanoscale technologies to enable a range of therapies for organ failure, cardiovascular disease and cancer. In enabling this vision he works closely with clinicians (including interventional radiologists, cardiologists and surgeons). For example, he has developed numerous techniques in controlling the behavior of patient-derived cells to engineer artificial tissues and cell-based therapies. His group also aims to engineer tissue regenerative therapeutics using water-containing polymer networks called hydrogels that can regulate cell behavior. Specifically, he has developed photo-crosslinkable hybrid hydrogels that combine natural biomolecules with nanoparticles to regulate the chemical, biological, mechanical and electrical properties of gels. These functional scaffolds induce the differentiation of stem cells to desired cell types and direct the formation of vascularized heart or bone tissues. Since tissue function is highly dependent on architecture, he has also used microfabrication methods, such as microfluidics, photolithography, bioprinting, and molding, to regulate the architecture of these materials. He has employed these strategies to generate miniaturized tissues. To create tissue complexity, he has also developed directed assembly techniques to compile small tissue modules into larger constructs. It is anticipated that such approaches will lead to the development of next-generation regenerative therapeutics and biomedical devices.

Aditya Kunjapur, Thomas Willing Early Career Associate Professor of Chemical & Biomolecular Engineering, University of Delaware, United States of America

Engineering Microbes to Create and Harness Non-Standard Amino Acids

In this talk, I will describe my group’s efforts to genetically engineer microbes that can harness new-to-nature building blocks, specifically non-standard L-alpha amino acids (nsAAs). Besides the typical work advanced by the genetic code expansion that allows ribosomal translation of proteins that contain these nsAAs, I will discuss how it can be advantageous in numerous contexts for a microbe to be engineered to biosynthesize the nsAA either fully or partially. Lastly, I will describe how microbes can be engineered to rely on nsAAs for their survival. Together, the use of nsAAs in polypeptide-based materials or the use of nsAAs to provide spatiotemporal control of engineered microbial survival have promise for biofabrication and biomanufacturing, particularly in biomedical or agricultural applications.

​

Mid-Afternoon Coffee Break and Networking in the Exhibit Hall

​

Kristopher Kilian, Professor, University of New South Wales, Sydney Australia

Biomaterials Design for Biofabrication and Biomanufacturing

Most hydrogels used in biomedical applications display a homogenous static architecture. In contrast, natural hydrogels in tissue are highly dynamic, where internal and external forces will catalyse changes in chemistry, architecture, and mechanical properties. In this presentation, I will discuss how materials chemistry can be used to fabricate dynamic hydrogels that mimic signalling in natural tissue. First, I will show how supramolecular assembly can be used to create hierarchically structured hydrogels with tuneable mechanics and self-healing behaviour. Next, I will demonstrate how modifying the biochemical and biophysical attributes can promote desirable activities in stem cells, towards next-gen support matrices for cell production. Finally, the potential of using these soft materials as cell delivery vehicles will be presented, with examples of tailored release through integrating complementary assembly motifs. Together, these examples demonstrate how the principles of self-assembly and biorecognition serve as powerful drivers in materials design for biotechnology and biomedical applications.

Carolyn Mills, Assistant Professor, Department of Bioengineering, University of California-Santa Barbara, United States of America

Developing an Anaerobic Cell-Free Protein Expression Platform for Discovery of Novel Enzyme Function

From the human gut to the soils that support plant growth, many of the microbes that support life thrive in anaerobic environments. These microbes often thrive in these environments by producing proteins that facilitate unique chemical transformations; notable examples include lignocellulosic biomass degradation and nitrogen fixation. To date, however, our ability to harness and engineer these functionalities has been stymied by challenges associated with mapping gene to function relationships in anaerobic organisms. These include a lack of genetic tools for non-model, anaerobic organisms and difficulties with heterologous expression systems (e.g., limited access to relevant post-translational modifications or maintenance of anaerobic conditions for oxygen-sensitive proteins). To address these challenges, we propose the development of anaerobic cell-free protein synthesis (CFPS) systems derived from these anaerobic organisms.

In this talk, I will describe I work towards this goal. First, we have identified fluorescent proteins compatible with anaerobic conditions—iRFP702 and mFAP2a—that can serve as reporters of protein expression in anaerobic cell-free expression experiments. We find that both reporters function aerobically and anaerobically in CFPS, producing good signal above background, in contrast to an sfGFP control, which only produces signal in the presence of oxygen. To enable future high-throughput screening of genes of interest in the future, we have also investigated miniaturization of CFPS reactions using acoustic liquid handling. Experiments with our two fluorescent reporters—iRFP702 and mFAP2a—as well as the sfGFP reporter commonly used for optimizing CFPS indicate that total reaction volume can be scaled down to 0.5 µL while still maintaining adequate signal-to-noise for all three reporters. This work established an accurate and reproducible liquid dispensing method for future exploratory experiments and will allow for fully automated workflows. Ultimately, we envision coupling this workflow with other automated processes in the NSF Biofoundry for Extreme and Exceptional Fungi, Archaea and Bacteria (ExFAB) housed at UC Santa Barbara to efficiently optimize CFPS in lysates derived from non-model organisms and, eventually, screen genes of unknown function. These developments will expand our ability to study these strains while work continues for the development of genetic tools for strain development.

Genan Wang, Researcher, North Carolina State University, United States of America

Engineering Probiotic Yeast as a Live Biotherapeutic Platform for Vitamin Delivery to the Gut

Live biotherapeutic products (LBPs) can improve host health by delivering therapeutics or micronutrients to specific areas of the body. In our study, we used the probiotic yeast Saccharomyces boulardii to synthesize vitamins A, C, and E in proportion to human nutritional needs. We utilized metabolic engineering and expression-level tuning to enhance vitamin yields. Our long-term vision is to develop these strains as either a food additive or a living probiotic supplement capable of producing vitamins in vivo.