Hybridoma Services

• 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

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

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-HOCK^® (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)