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-HOCKTM (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 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®) -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 Screeners enable 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® (SPR) and Octet®, recombinant production at any desired scale, humanization/affinity maturation if needed)