Cell-Based Assays

Curia performs a wide variety of cell-based assays to support drug discovery efforts in both the small molecule and large molecule arenas. Our Assay team has a broad experience base in culture and implementation of standard cell lines, iPSCs, primary cell lines and PBMCs utilizing a wide variety of readouts and technologies. The assay team leverages the full menu of Multimode Readers, Cell Imaging/Analysis instrumentation, Genetic Assay Technologies and Analytical Instruments available across Curia. We maintain BSL2+ level cell culture labs and utilize segregation and isolation of Tissue preparation, Viral workflows as well as a quarantine for new cell lines received from clients and collaborators. A variety of liquid handling technologies and platforms are available to support studies at a small scale as well as at full HTS scale

Assay Type Highlights:

• Standardized Assays: ADCC/CDC, T-Cell Functional Assays, Viral Ab Neutralization, TCID50 and Cytotoxicity…fine tuned to client need
• Cell stimulation workflows with multiple readouts
• Reporter Gene/Immune Checkpoint assays
• PBMC-based workflows: Commercial Thaw-and-Use cells or fresh via on-site Research Blood Donor program
• Custom Bioassay Cell Line generation, RCB and MCB workflows
• Full customization of workflow and assay formats available

Assay Format/Technology Highlights:

• Fluorescent/Luminescence: HTRF, FP, FRET, AlphaLISA, AlphaScreen, Glo, Luciferase
• Flow Cytometry and FACS
• ELISA-Based: Meso Scale Discovery ECL and traditional TMB
• cIEF/CE-SDS and Traditional PAGE/Western (ECL and Fluorescence Imaging)
• Epi-fluorescence Microscopy, Confocal and High Content Screening (HCS)
• Spectrophotometric Abs and Radiometric Readouts
• Full Custom Development; Discovery through cGxP Assay Support

Liquid Handling Highlights:

• Formulatrix Tempest Automated Dispenser Platform
• 96/384 Channel Pipettors from Integra Biosciences and Apricot Technologies
• Tecan d300e Digital Dispenser
• Labcyte Echo 550 (3rd Party Collaboration)

Cell-Based Assay Case Study:

Antibody Neutralization Assay in HEK293 Cells

Purpose: Recurrent flow cytometry screening of non-human primate serum samples for neutralizing antibodies to a particular set of viruses in support of Client’s Rare Disease Unit

Design: HEK293 cells are transduced with MOI of 10e4 virus particles either neat or pre-incubated with heat-inactivated test serum samples in titration for 48 hours to determine neutralization values of transduction. Readout by flow cytometry to detect virally expressed GFP. Wt virus capable of 100% transduction @ 10e4 MOI; Mutant-only capable of ~7-14% transduction @ 10e4 MOI

Optimization Work: Confirmation of functionality of Wt and Mutant viruses and establish consistency with existing literature data for transduction levels. Scaling of workflow for maximum sample throughput

Outcome: Currently in recurrent support modality; batches of 50-150 samples. Additional viral variants optimized and in testing