Hit Discovery

Goal: Identification of unique antibodies that are specific for your target with broad epitope coverage.

Principle: For each target structure, up to hundreds of different, antigen-specific antibodies are present in our fully human antibody library (depending on type, nature and size of the antigen). These antibodies can be selected, isolated and analyzed based on their binding characteristics in-vitro. As a result, we can identify antibodies that display:

  • pH or salt sensitive target binding
  • no cross-reactivity to homologous proteins
  • high epitope specificity and selectivity
  • species x-reactivity

Application: Generation of tool antibodies, identification of potential therapeutic candidates.

The antibody Hit Discovery technology includes the following modules that can be combined individually:

hit-discovery-grafic

Antibody target preparation

  • High-quality peptide synthesis
  • Recombinant protein production, supply, modification and/or QC
  • Transient overexpression of target genes on cells
  • Stable target cell line development

Antibody library

  • Universal fully human antibody library ready to start
    (nature-derived sequences, >1011 total diversity, germline specific sublibraries available or as pool)
  • Custom libraries can be generated from human patients or immunized mice, rabbits, goats or llamas

Antibody selection

In-vitro selection:

  • on surface immobilized antigens
  • on antigens in solution
  • on whole cells

Control of binding conditions:

  • pH, salt, or buffer conditions
  • competitions with ligand or other antibodies for epitope specific antibodies
  • negative selection with isoform or homologous antigen to avoid cross-reactivity
  • alternating panning with antigen of different species for interspecies cross-reactivity

Antibody screening

  • Production of soluble antibodies instead of antibody-phage particles
  • ELISA on up to 4 different antigens in a single run
  • Flow cytometry on cells (up to 3 cell lines in a single run)
  • Off-rate ranking of antibodies (BLI)

Antibody sequence analysis

DNA sequencing of antigen specific antibodies & bioinformatic analysis:

  • Framework and CDR analysis and annotation
  • Identification of potential PTMs in CDRs
  • Modelling of biochemical properties and developability ranking (e.g. surface and CDR hydrophobicity/charge, pI, germinality index, unusual amino acid residues)

Antibody reformatting and production

  • Reformatting into the mono- or bivalent antibody format (e.g. scFv, Fab, scFv-Fc, IgG; each format with different isoforms, Fc species or tags available).
  • Production by transient transfection of mammalian cells (typically HEK293, CHO on request)
  • Scalable up to 100 mg for selected lead candidate(s)
  • Low endotoxin available (< 1 EU/mg)
  • Stable cell line generation with external partner

Validation and analytics

  • ELISA
  • Flow cytometry
  • Affinity measurement (Kinexa, BLI)
  • Gelfiltration/Size exclusion chromatography (with/without stress conditions)
  • Immunofluorescence
  • Immunohistochemistry
  • Functional bioassays (assay dependent, on request)

YUMAB® Antibody Discovery

  • Fully human, nature-derived antibody sequences
  • Hapten, peptide, proteins and whole cells as target antigen
  • Pre-designed antigen binding by controlled in vitro selection

Contact

YUMAB GMBH
Science Campus Braunschweig-Süd
Inhoffenstr. 7
38124 Braunschweig – Germany

Email: info@yumab.com
Phone: +49 531 481170-0