Variable Region Sequencing
All antibodies have the same basic protein structure but exhibit sequence variability in those regions that contact the antigen. These so-called “variable regions” represent an approximately 100 amino acid domain found at the amino terminals of both the light and heavy chains of IgG. Different monoclonal antibodies to the same antigen target can also have different variable region sequences. Because hybridomas synthesize antibody molecules with identical antigen specificity, they are an ideal source of genetic material for the preparation of recombinant molecules.
- Hybridoma Identification
- Antibody Engineering
- Antibody Humanization
Our approach uses mRNA isolated from actively growing hybridoma cells with light and heavy chain primers for sequencing. Cloning of PCR amplified variable region cDNA into appropriate vectors permits further engineering of recombinant molecules. You can confirm your cDNA sequences by comparing them with the protein sequence of the purified antibody.