Antibody Sequencing/ Recombinant Antibody Production (NB-SC110)
Novatein Biosciences provides comprehensive monoclonal antibody cloning and antibody sequencing service: The antibody (IgG, IgM, IgA, or IgE) variable regions of the heavy chain (VH) and the light chain (VL) from mouse, rat, hamster, rabbit, monkey, or human hybridomas or clonal B cells are PCR amplified and sequenced. To be 100% sure about the results of PCR-based hybridoma antibody cloning and antibody sequencing, antigen-antibody binding is confirmed by transiently expressing recombinant single-chain antibodies (scFv) or whole IgGs in HEK 293 or CHO cells, and testing whether the recombinant scFv or whole IgG retains the antigen binding affinity and specificity.
100000 cells per hybridoma are sufficient for cloning and sequencing of antibody genes.
General procedure:
1) RNA extraction from hybridoma or clonal B cells;
2) cDNA synthesis
3) 5’ RACE amplification using constant regions of VH and VL
4) Cloning of PCR fragments
5) Sequencing
7) CDR analysis
Turnaround: 2-3 weeks