De Novo Sequencing
During the discovery process of therapeutic antibodies, many important monoclonal antibodies reagents can be used as positive controls for in vitro or in vivo experiments. However, the hybridoma cell line for most of the reagents are typically not available and there is often the need for obtaining VH and VL sequences for expression or engineering of the molecule. De novo sequencing of antibody by mass spectrometry is a technology that can analyze unknown antibodies and retrieve full VH and VL sequences for a given antibody. Briefly, the antibody protein (~0.1 mg or more total protein needed) is digested with multiple site specific or semi-specific proteases, the peptides are separated by liquid chromatography and analyzed by thermo orbitrap mass spectrometry for de novo peptide sequencing. The overlapping peptides are assembled by bioinformatics tools to obtain a fully integrated VH or VL sequence.
We will deliver a functional antibody sequence along with a comprehensive sequencing report with full sequence coverage. The sequence can be validated by the client for recombinant antibody expression followed by binding/functional analysis of the antibody.