Antibody generation, characterization and optimization

We have experience generating antibodies against hundreds of different antigens, including:

• G protein–coupled receptors (GPCRs)
• Immunomodulators
• Ion channels
• Transporters
• Kinases, phosphatases and other enzymes
• Antibody idiotypes
• Protein complexes
• Histones
• Peptides
• Glycans
• Small molecules

• Libraries:
  • Naïve libraries from camels, alpacas, and llamas: selection of sdAbs (VHHs) against non-immunizable targets
  • Immune libraries from llamas: protein immunization, DNA immunization, whole cell immunization, multiplexed / batch immunization
  • Large synthetic human sdAb libraries: variable heavy (VH) and variable light (VL) sdAbs with engineered scaffolds (size: >10¹⁰)
  • Consistent isolation of sdAbs from immune libraries in low nM – pM affinity range
• Screening and panning:
  • Panning for cell-surface molecule binding sdAbs, cell internalizing sdAbs, cross-species and isoform-specific sdAbs, high affinity binders, binders with high stability, pH-specific binding, and binders to a specific epitope (competitive elution)
  • Panning against immobilized proteins, protein-domains, and immobilized proteins in specific orientation
  • Next-generation sequencing (NGS) to identify binders

• Protein immunization, DNA immunization, whole-cell immunization, multiplexed / batch immunization
• Immunization of mice and rats
• Genetic immunization for complex proteins: single-pass and multiple-pass transmembrane proteins, and other proteins
• Panning for cell-surface molecule binding antibodies, cell internalizing antibodies, cross-species and isoform-specific antibodies
• Panning against immobilized proteins, protein-domains, and immobilized proteins in specific orientation
• High-throughput hybridoma generation: multiplexed immunization, electrofusion, secretor cloning and clone picking

• Biophysical characterization:
  • Melting temperature (Tm)
  • Long-term stability
  • Aggregation
  • Secondary and tertiary structure
  • Refoldability
  • Sequencing
  • Immunoglobulin (Ig) subclass determination
  • Kinetics and affinity
  • Epitope binning / mapping
• In vitro functional screening:
  • High-throughput cell binding
  • High-throughput cell internalization
  • Enzyme inhibition
  • Ligand competition / blockade
• Computational characterization:
  • Antibody structure modeling
  • Binding affinity analysis
  • Protein-protein docking
  • Molecular dynamics simulations
  • Protein electrostatics

• Engineering:
  • Stability improvement
  • Affinity improvement
  • Incorporation of tags for labeling or conjugation
  • Generation of multi-valent and multi-specific sdAb formats
  • Half-life extension using anti-serum albumin sdAbs
• Molecular modeling and design:
  • Humanization for reduced immunogenicity
  • Virtual affinity maturation: Assisted Design of Antibody and Protein Therapeutics (ADAPT) platform
  • Computational developability assessment: stability, immunogenicity, aggregation

• Mirrorball microplate cytometer
• Clonepix FL mammalian cell clone picker
• FACS Aria II cell sorter
• Surface plasmon resonance (SPR) instruments: Biacore T200s and Biacore 3000s
• Isothermal titration calorimetry (ITC)
• Circular dichroism spectroscopy (CDS)
• Size exclusion chromatography (SEC)
• Plate washers, dispensers, readers
• KingFisher purification system
• BSL-2 laboratories

Efficacy and safety of therapeutic antibody candidates can be validated through our functional characterization and analytics expertise, and at our preclinical in vivo facility. Production of antibodies is offered by NRC's Human Health Therapeutics biomanufacturing experts, with scale-up at our microbial fermentation pilot plant and cell culture pilot plant.