20000 cells per hybridoma clone are sufficient for cloning and sequencing the monoclonal antibody variable regions (both VH and VL) and full length. Welcome to challenge us with smaller number of clonal B cells or non-viable cells for hybridoma rescue.
Both Sanger sequencing-based and nanopore sequencing-based methods are available for antibody sequencing from hybridoma and clonal B cells. Sanger sequencing is labor intensive and time consuming so that it is difficult to reduce the cost for a large number of samples. However, nanopore sequencing can be used to reduce the cost signicantly, especially for a large number of samples.
De novo antibody sequencing to obtain the intact amino acid sequence of variable and/or constant regions of monoclonal antibodies by LC-MS/MS.
Sample requirement: 3-5 mg at 1 mg/ml preferred (minimum: 2 mg at 1 mg/mL).
Fast turnaround time: 2-3 months per antibody, but may require more time depending on the complexity of analysis.Single B cell antibody sequencing has become popular for antibody discovery, especially for those difficult to obtain. After antigen specific single B cell sorting, the single B cells can be directly used for antibody seqencing or cultured for 2 weeks before antibody sequencing. Single B cell sequencing by next generation sequencing (NGS) including nanopore sequencing has been successfully applied to single B cell antibody technologies.