EMU: Beacon-high throughput screening of single cells by optofluidics

• Grant number: 22/11436-3
• Support Opportunities: Research Infrastructure Program - Technology and Innovation
• Start date: June 01, 2023
• End date: May 31, 2030
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Ana Maria Moro
• Grantee: Ana Maria Moro
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Esper Georges Kallás ; Irina Kerkis ; Roxane Maria Fontes Piazza
• Associated scholarship(s): 24/15901-8 - B cells immunophenotyping, PCR, sequencing analysis, and genes cloning for expression of monoclonal antibodies, BP.TT; 24/13757-7 - Chromatography and analytical tecniques for monoclonal antibodies (structure and function), BP.TT

Abstract

The development of biopharmaceuticals involves several complex steps that demand a lot of time and resources, in addition to the risks of failure. When working with monoclonal antibodies (mAbs), the unpredictability factor is high because it depends on finding, among the peripheral blood B cells, or among the B cells of organs of immunized animals, the only ones that express the antibody(s)( s) of interest. A recent example can be given by the anti-SARS-CoV-2 mAbs, which are only relevant in therapeutics when they are highly neutralizing, have adequate pharmacokinetics, do not cause ADE (antibody dependent enhancement) and can bind and neutralize the variants. It is about looking for a "needle in a haystack" because these antibodies are rare. The same goes for finding the phenotypically relevant cells for cell therapy development. The possibility of performing initial screening based on functional studies of binding and/or recognition of other cells or molecules can make a big difference in finding the most important cells. The possibility of eliminating cells in apoptosis, performing multiparametric analyzes on 100,000 cells in real time opens a universe for the development of relevant biopharmaceuticals. Once the gene sequences of interest are found, it is necessary to transfect them into host cells for stable expression. Again, a random process occurs, and to adequate the process to the possible conditions, the number of cells tested is small and does not represent the universe of variability that allows finding the best clone manually. The Beacon equipment is also very suitable for the development of cell lines, when the growth performance and quality of the antibody are analyzed inside the microchips before moving on to the scaling step. The Beacon works as a cell sorter, which isolates cells through real-time functional analysis, operates in a sterile environment (by negative pressure), incubates cells in a CO2 environment and allows cell growth by perfusion of the culture medium in nanopens. The selected clones are exported inside the equipment to culture plates for the continuity of the process. We and our associated partners have several ongoing projects that can benefit from the use of Beacon and we have other innovative projects in incubation that would only be possible through this platform. (AU)