How we work

Generation of monoclonal hybridoma cell lines is divided into three phases:

I) Immunization and cell fusion

II) Screening, clone expansion, cryoconservation, IgG subclass determination

III) Subcloning

 

Antigen Selection

  • proteins, peptides, exosomes, VLPs and other antigens

 

Immunization and Fusion

  • Immunization of rats and mice
  • Fusion of spleen B cells with a HAT-sensitive mouse myeloma cell line

 

Screening and validation

  • Primary high-throughput screening of hybridoma supernatants
  • Expansion and cryoconservation of positive oligoclones
  • Determination of titer and IgG subclass
  • Further validation by collaboration partner

 

Subcloning

  • Single cell cloning of selected hybridomas to create stable monoclonal hybridoma
  • Cryoconservation for long-term viable storage in liquid nitrogen

 

Antibody Production

  • Expansion of hybridoma cells and collection of antibody supernatants

 

Antibody Purification and Labeling

  • Protein A / Protein G affinity chromatography
  • Gelfiltration
  • Endotoxin removal
  • Conjugation with HRP, Biotin or fluorescent dyes