Nanoscale integration of single cell biologics discovery processes using optofluidic manipulation and monitoring

Abstract

The new and rapid advancement in the complexity of biologics drug discovery has been driven by a deeper understanding of biological systems combined with innovative new therapeutic modalities, paving the way to breakthrough therapies for previously intractable diseases. These exciting times in biomedical innovation require the development of novel technologies to facilitate the sophisticated, multifaceted, high‐paced workflows necessary to support modern large molecule drug discovery. A high‐level aspiration is a true integration of “lab‐on‐a‐chip” methods that vastly miniaturize cellulmical experiments could transform the speed, cost, and success of multiple workstreams in biologics development. Several microscale bioprocess technologies have been established that incrementally address these needs, yet each is inflexibly designed for a very specific process thus limiting an integrated holistic application. A more fully integrated nanoscale approach that incorporates manipulation, culture, analytics, and traceable digital record keeping of thousands of single cells in a relevant nanoenvironment would be a transformative technology capable of keeping pace with today's rapid and complex drug discovery demands. The recent advent of optical manipulation of cells using light‐induced electrokinetics with micro‐ and nanoscale cell culture is poised to revolutionize both fundamental and applied biological research. In this review, we summarize the current state of the art for optical manipulation techniques and discuss emerging biological applications of this technology. In particular, we focus on promising prospects for drug discovery workflows, including antibody discovery, bioassay development, antibody engineering, and cell line development, which are enabled by the automation and industrialization of an integrated optoelectronic single‐cell manipulation and culture platform. Continued development of such platforms will be well positioned to overcome many of the challenges currently associated with fragmented, low‐throughput bioprocess workflows in biopharma and life science research.

Dynamic single-cell phenotyping of immune cells using the microfluidic platform DropMap

“…They do not allow the quantitative characterization of secreted proteins by living cells. Recently, commercially available technologies based on compartmentalizing cells in microfabricated chambers have been developed to perform a range of assays, including real-time IgG and cytokine secretion, apoptosis and cell-cell interaction [39][40][41] . For example, the Berkeley Lights system allows optofluidic handling and analysis of single cells, including recovery of individual cells for sequence analysis, but can analyze ~10,000 cells as the system currently has 14115 chambers ("nanopens"), whereas the DropMap system can currently analyze up to 300,000 cells.…”

“…xii) Immediately proceed to image the droplets as described in steps [38][39][40][41][42][43][44][45] Step 39, use the following image acquisition settings: DAPI (300 ms), GFP (200 ms), CY3 (200 ms), CY5 (200 ms) and BF (20 ms) images using a 10X objective. 100 fields of view (10x10) every 10 minutes during 60 min (7 time points).…”

Single B cell technologies for monoclonal antibody discovery

“…Here, secreted antibody binding to the antigen-coated beads is detected by the secondary fluorescent anti-IgG antibody, generating a fluorescent bloom. The screening phase takes usually only a few minutes to up to several hours, followed by specific single B cell retrieval using a structured light-based system termed Opto Electro Positioning (OEP) [73,74].…”

Membrane protein production and formulation for drug discovery

“…Although binding can be detected by various methods [e.g., fluorescence-activated cell sorting (FACS) and ELISA], selecting mAbs with inhibition or activation activity necessitates more discriminating functional assays. Cutting-edge single-cell sorting methods (i.e., Beacon optofluidics platform ix ) promise to allow binding and functional screening to occur in parallel, greatly accelerating the mAb discovery timeline [84]. Another innovative functional screen was recently developed whereby single-domain antibodies were anchored to the cell membrane together with the apelin receptor GPCR target and downstream signaling (β-arrestin recruitment) was monitored via FACS assay.…”