Live SARS-CoV-2 Research: Interlocking Pieces Working Together
Boston University (BU), National Emerging Infectious Diseases Laboratories (NEIDL)
When Dr. Robert Davey started researching Ebola virus drug development at the University of Texas Medical Branch (UTMB) at Galveston back in 2008, he had no idea just how keenly his work would prepare him to play a pivotal role in researching treatments for a very different global pandemic. In fact, a number of pieces fit together to support his current role in screening small molecule compounds that may be used in the fight against SARS-CoV-2, the novel coronavirus that causes COVID-19.
The Process Fits
When Dr. Davey moved from UTMB to the Texas Biomedical Research Institute in San Antonio, he developed a pipeline for effectively screening small molecule drug compounds in a biosafety level four (BSL-4) maximum containment laboratory.
Given the rigorous safety and security measures in a BSL-4 laboratory, the lab was equipped with small automated products, including many from BioTek Instruments, that could improve throughput while reducing manual interactions, reduce user safety risks, and increase accuracy. If something happened to these tools, they could be decontaminated and easily removed from the lab, unlike large robotic systems that would require the services of a highly specialized technician.
With a process in place, Dr. Davey set up a new lab at the Boston University’s (BU) National Emerging Infectious Diseases Laboratories (NEIDL) in 2018. As the SARS-CoV-2 outbreak spread in late 2019, he realized that his drug screening process at high containment would work very well for the novel coronavirus, and quickly pivoted his research efforts towards that goal.
At National Emerging Infectious Diseases Laboratories (NEIDL) March 20, 2020, member of the Davey Lab include Callie Donahue, PhD student, from left, Hiroyuki More, DVM, PhD, Robert Davey, PhD, and Manu Anantpadma, PhD. The team’s goal is to develop a small molecule drug treatment to stop infection of SARS-CoV-2.
Photo by Cydney Scott for Boston University Photography
Welcome to the BioTek Universe
“We need flexibility and modular, interlocking interaction without the hassles of reprogramming everything, and that’s why we created a ‘BioTek Universe’ within our lab here at Boston University,” he remarks. This so-called Universe consists of two Cytation™ 1 Cell Imaging Multi-Mode Readers, two BioStack™ Microplate Stackers, and two MultiFlo™ FX Multi-Mode Dispensers, and is expected to soon expand to include a third BioStack and a 405™ LS Microplate Washer.
Scores of 384-well microplates containing cells treated with drug compounds, SARS-CoV-2, and stained with fluorescently-labeled antibodies or oligonucleotides are loaded onto the BioStacks each evening. Overnight, the BioStacks feed plates into the Cytations for high-content imaging. Dr. Davey appreciates that the Cytations have a small footprint in a lab where space is highly limited, an autofocus feature, and barcode reading through the stackers that integrates well with the software to track samples and strengthen quality control.
Dr. Davey also makes use of Cytation’s 16-bit camera, pointing out that, “A sensitive 16-bit camera provides 65,000 shades of gray, and for me, that means more breadth of signal in the assay, compared to the more common 12-bit camera with about 4,000 shades of gray.”
Photo by Cydney Scott for Boston University Photography
Creating a Robust Foundation
At the onset of this coronavirus research project, Dr. Davey was able to obtain SARS-CoV-1 virus antibodies that cross-reacted with SARS-CoV-2. Cytation quickly identified poorly-responding antibodies that were then rejected. Additionally, Cytation was used to narrow the scope of host cells to those that were robust and amenable to high-throughput. “Consistency is absolutely critical to our process,” he says. “By having it work the same every time, and eliminating potential nuances, we can be confident that when there is a blip on the radar, so to speak, it’s really important.”
Putting It All Together
The biggest screen that Dr. Davey’s lab has completed to date included 28,000 data points, and the ‘BioTek Universe’ continues to perform reliably while offering flexibility. Dr. Davey likes knowing that they can move different BioTek elements around without disrupting the workflow or communication.
He notes that while BioTek’s Gen5™ software is very good, they use a different software platform for their analyses. “Other platforms incorporate a proprietary image format, and to me, that doesn’t make sense; it’s restrictive,” he comments. “BioTek uses an open image format so that we can export the images. This gives me the flexibility I need to get as much information as possible from the images.”
SARS-CoV-2 drug screening images taken using Cytation 1. 384-well plates were challenged with virus in the presence of small molecule treatment candidates. After 36 hours, cells were fixed in formalin, then stained with anti-N antibody followed by Alexa 448-labeled secondary antibody to detect infected cells (green). Cell nuclei were stained with Hoechst 33342 (blue). After imaging the entire plate, images were processed using high content image analysis software and cell nuclei and number of infected cells calculated. Images show (A) normal infection with many infected cells and (B) reduced infection efficiency in treated cells.
To learn more about Boston University (BU), National Emerging Infectious Diseases Laboratories (NEIDL), visit their web site.
Thanks to Dr. Robert Davey at the National Emerging Infectious Diseases Laboratories (NEIDL) for sharing his BioTek experience.