Cortalix offers custom nanobody services. Our services can be limited to just a single, small sub-project, such as the selection of a high-affinity binder for a target or a sandwich ELISA for that target. Or to a complete start-to-finish program to deliver a number of well-characterized, humanized, high-affinity antigen-binding nanobodies with an appropriate functional group to chelate a radionuclide of interest. Or everything in between. Cortalix strives to bring the same level of dedication to every project.

1. Phage Display, Panning and Selection

  • Access to an immune VHH library after llama immunization, cDNA preparation, and cloning into a phagemid vector, or,
  • Access to a synthetic VHH library by randomization of CDRs in a generic scaffold
  • Production of a VHH-phage library in E. coli
  • Biopanning to enrich for specific target binders using immobilized antigen or whole cells
  • DNA sequence analysis of high affinity binders
  • Further characterization of purified VHHs (ELISA, SPR, immunofluorescence, cells, tissues, in vivo disease models)

2. Cloning and production in E. coli and P. pastoris

  • Construction of expression vectors for VHH production in bacteria and/or yeast
  • Small scale production in E. coli (0.1 – 2 mg) for early characterization
  • Medium scale production in P. pastoris (2-50 mg) for further in vitro and in vivo characterization
  • Microbial cell bank preparation
  • Semi-large scale production (5-100 mg) and pre-GMP technical batch optimization, up to 7 l fermentation

3. Additional Services

  • Humanization and liability analysis
  • Spacer or linker design
  • Conjugation of functional groups (fluorescent or NIR dyes, NOTA/DOTA/DTPA, etc.)
  • Multispecific and paratopic nanobodies design
  • ELISA assay development
  • Inducible recombinant antigen expression in HEK293 for cell-based biopanning and characterization
  • FACS analysis
  • SPR analysis