Hybridoma Services
Curia’s in vivo Discovery Immunology team offers high-performance hybridoma-based technologies to help clients discover great antibodies that make a difference.
- A variety of proprietary immunization protocols (DNA, protein, peptide, small molecule, and cell-based) are available with demonstrated success in multiple wild-type murine strains and fully-human antibody-producing transgenic mice, with advanced tolerance-breaking approaches when needed (including use of specialized target-deficient mice).
- ELISA- and FACS-based high-throughput screening and off-rate analyses of hybridomas indicate the specificity, cross-reactivity, and functionality of candidate antibodies as early as two months from campaign initiation.
- Diverse therapeutic and clinical diagnostic/imaging candidates against immuno-oncology targets, GPCRs, neurobiology targets, enzymes, autoimmune/inflammatory targets, metabolic targets, and small molecules.
Discovery Immunology mAbs can seamlessly transition to downstream GXP production, enabling one-stop shopping from discovery to development.
Workflow of Hybridoma Discovery Immunology
| Upfront Due Dilligence | Immunizations | Hybridoma Screens | Antibody Characterization |
|---|---|---|---|
| – Target analysis and antigen design – Goal-oriented proposal development – Antigen biofunction QC | – Rapid immunization protocols – DNA, protien, peptide, and cell-based immunizations – In-life titer checks enable real-time optimization of immunizations, and selection of animals with optimal titers | – Hybridoma generation by electrofusion – 384-well plate high throughput screens (ELISA or multiplex FACS) – Data Master Files for candidate selection | – Hybridoma single cell cloning and variable region sequencing – mAB characterization affinity, binding EC50 (ELISA or FACS), biofunction IC50 or EC50 (customized functional assays such as ligand blocking and receptor internalization) |
Complete campaigns incorporate Curia’s unique suite of integrated services including:
- Target analysis and immunogen production
- Immunization, hybridoma generation, and single-cell cloning
- Variable region sequencing and purified mAb production/characterization
Key Highlights
Chain of Discovery™
- Rigorous upfront target analysis and goal-oriented proposal development
- Data updates provide tracking from target to mAb delivery
- Delivery of sequence-confirmed hybridomas with variable region sequences
Immunizations
- Multiple wild-type mouse or rat strains
- DNA, protein, peptide, and cell-based immunizations
- Often take multiple immunization approaches against a single target to generate a large diverse repertoire of candidate antibodies.
Curia’s advanced tolerance-breaking approaches can open epitopic space for targets that are highly conserved between humans and mice. They can also increase opportunities for discovering cross-reactive antibodies or for generating surrogate anti-mouse target antibodies.
Characterization
- Analysis of heavy and light chain V- and J- gene usage and of CDR3 sequence
- 384-well high-throughput ELISA or FACS screening of thousands of hybridomas for target binding
- Incorporation of cell-based ligand blocking assays
- ELISA, BLI, Carterra Arrayed SPR epitope binning and affinity determination
Proprietary PentaMice® Platform for Maximum Plasma Titers
- Royalty-free set of wildtype mice designed for in vivo immunizations to achieve maximum plasma titers
- The platform consists of 5 WT strains that cover 9 distinct MHC haplotypes
- 10 mice total (2 mice per strain) are included
Download a free eBook: Introduction to Hybridoma Technologies for Antibody Discovery
Download our Whitepaper: PentaMice® platform for COVID-19 antibody discovery
Other differentiators:
- Multiple immunization strategies (protein, DNA, cell-based immunogens) using a variety of WT strains and/or human-antibody producing transgenics and/or target-specific KOs coupled with in-life plasma titer checks allows us to track antibody responses in vivo and focus on the most promising animals for subsequent fusion and antibody recovery.
- Antibody Discovery at Curia typical begins with rodent immunizations. For most protein or peptide immunogens, Curia utilizes our highly effective HT-HOCKSM (High Titer Hock) immunization platform. HT-HOCK immunizations use proprietary combinations of adjuvants and optimized injection intervals to quickly generate in vivo plasma titers beyond industry and academic norms. In-life plasma titer checks begin on day 17 and are performed weekly throughout the HT-HOCK immunization process. This enables monitoring and fine-tuning of in vivo antibody responses, resulting in high plasma titers and superior antibody discovery hit rates. Whether it’s hybridoma-, phage or yeast display-, or single B cell (Bruker® Beacon® Optofluidic System) -based antibody discovery, Curia’s HT-HOCK immunization platform delivers maximum antibody discovery success.
- Expertise in innovative tolerance-breaking approaches for targets with high human-mouse identity or if human/mouse target cross-reactivity is required.
- 384-well screening format enables high capacity enables interrogation of 4,000-12,000 hybridomas in a standard campaign.
- Intellicyt® iQue® Screener PLUS platform enables high-throughput multiplex flow cytometry-based primary screening to immediately identify hits with desired specificity and cross-reactivity requirements.
- Functional assay development and integration into screening cascade.
- Seamless integration with Curia’s complete range of analytic and production capabilities (immunogen production, cell line generation, hybridoma sequencing, purified antibody production, affinity measurements by Carterra® LSA® platform (SPR) and Octet® system, recombinant production at any desired scale, humanization/affinity maturation if needed)