@article{hetzler_marinakos_lott_mohammad_lass-napiorkowska_kolbe_turrentine_fields_overton_marie_et al._2024, title={Adeno-associated virus genome quantification with amplification-free CRISPR-Cas12a}, volume={3}, ISSN={["1476-5462"]}, DOI={10.1038/s41434-024-00449-x}, journal={GENE THERAPY}, author={Hetzler, Zach and Marinakos, Stella M. and Lott, Noah and Mohammad, Noor and Lass-Napiorkowska, Agnieszka and Kolbe, Jenna and Turrentine, Lauren and Fields, Delaney and Overton, Laurie and Marie, Helena and et al.}, year={2024}, month={Mar} }
 @misc{overton_boi_shastry_smith-moore_balchunas_sambandan_gilleskie_2023, title={Development and Delivery of a Hands-On Short Course in Adeno-Associated Virus Manufacturing to Support Growing Workforce Needs in Gene Therapy}, volume={34}, ISSN={["1557-7422"]}, url={http://dx.doi.org/10.1089/hum.2022.235}, DOI={10.1089/hum.2022.235}, abstractNote={The manufacturing of gene therapy products is a rapidly growing industry bolstered by the tremendous potential of these therapies to provide lifesaving treatment for rare and complex genetic diseases. The industry's steep rise has resulted in a high demand for skilled staff required to manufacture gene therapy products of the expected high quality. To address this skill shortage, more opportunities for education and training in all aspects of gene therapy manufacturing are needed. The Biomanufacturing Training and Education Center (BTEC) at the North Carolina State University (NC State) has developed and delivered (and continues to deliver) a four-day, hands-on course titled Hands-On cGMP Biomanufacturing of Vectors for Gene Therapy. The course, which consists of 60% hands-on laboratory activities and 40% lectures, aims to provide a comprehensive understanding of the gene therapy production process, from vial thaw through the final formulation step, and analytical testing. This paper discusses the design of the course, the backgrounds of the nearly 80 students who have participated in the seven offerings held since March 2019, and feedback from the course participants.}, number={7-8}, journal={HUMAN GENE THERAPY}, publisher={Mary Ann Liebert Inc}, author={Overton, Laurie and Boi, Cristiana and Shastry, Shriarjun and Smith-Moore, Caroline and Balchunas, John and Sambandan, Deepa and Gilleskie, Gary}, year={2023}, month={Apr}, pages={259–272} }
 @article{duffy_overton_flickinger_2022, title={A concept for continuous virus manufacture using a moving bed bioreactor: Growth of MDCK cells to confluence on paper as a model support}, volume={170}, ISSN={["1873-3204"]}, DOI={10.1016/j.cep.2021.108667}, abstractNote={A moving-bed bioreactor (MBB) could intensify growth of adherent mammalian cells for viral vaccines. A continuously fed sterile paper passing through a thin liquid layer as a flexible cell substrate is a new concept for bioprocess intensification (BPI). Paper could enable cell expansion with reduced footprint and reduced media consumption as the first stage of a 2-stage growth + infection continuous process. This study focused only on cell growth (stage 1). We report a simple 32 mm paper disc method to evaluate growth of adherent Madin Darby canine kidney cells (MDCK CCL-34) adapted to 5% FBS to confluence on unmodified papers to screen substrates. Bibulous paper was found to be the best substrate for proliferation of MDCK. An MBB process was simulated using paper discs to test growth to confluence (stage 1). Growth was characterized using staining, image analysis; confocal microscopy. Cells grew on the surface of bibulous paper to a confluence of >90% in 192 h. Extending this concept by stabilizing MDCK on paper (end of stage 1) by engineering cells to survive freezing or lyoprotection would enable live cells to be shipped as modules to multiple manufacturing sites for infection resulting in rapid, reproducible viral vaccine manufacture.}, journal={CHEMICAL ENGINEERING AND PROCESSING-PROCESS INTENSIFICATION}, author={Duffy, Colleen M. and Overton, Laurie and Flickinger, Michael C.}, year={2022}, month={Jan} }
 @article{hetzler_wang_krafft_jamalzadegan_overton_kudenov_ligler_wei_2022, title={Flexible sensor patch for continuous carbon dioxide monitoring}, volume={10}, ISSN={["2296-2646"]}, DOI={10.3389/fchem.2022.983523}, abstractNote={Monitoring and measurement of carbon dioxide (CO2) is critical for many fields. The gold standard CO2 sensor, the Severinghaus electrode, has remained unchanged for decades. In recent years, many other CO2 sensor formats, such as detection based upon pH-sensitive dyes, have been demonstrated, opening the door for relatively simple optical detection schemes. However, a majority of these optochemical sensors require complex sensor preparation steps and are difficult to control and repeatably execute. Here, we report a facile CO2 sensor generation method that suffers from none of the typical fabrication issues. The method described here utilizes polydimethylsiloxane (PDMS) as the flexible sensor matrix and 1-hydroxypyrene-3,6,8-trisulfonate (HPTS), a pH-sensitive dye, as the sensing material. HPTS, a base (NaOH), and glycerol are loaded as dense droplets into a thin PDMS layer which is subsequently cured around the droplet. The fabrication process does not require prior knowledge in chemistry or device fabrication and can be completed as quickly as PDMS cures (∼2 h). We demonstrate the application of this thin-patch sensor for in-line CO2 quantification in cell culture media. To this end, we optimized the sensing composition and quantified CO2 in the range of 0–20 kPa. A standard curve was generated with high fidelity (R2 = 0.998) along with an analytical resolution of 0.5 kPa (3.7 mm Hg). Additionally, the sensor is fully autoclavable for applications requiring sterility and has a long working lifetime. This flexible, simple-to-manufacture sensor has a myriad of potential applications and represents a new, straightforward means for optical carbon dioxide measurement.}, journal={FRONTIERS IN CHEMISTRY}, author={Hetzler, Zach and Wang, Yan and Krafft, Danny and Jamalzadegan, Sina and Overton, Laurie and Kudenov, Michael W. and Ligler, Frances S. and Wei, Qingshan}, year={2022}, month={Sep} }