Characterization of Unique Nanobodies by Native Mass Spectrometry
EMSL Project ID
60522
Abstract
Nanobodies contain the variable antigen binding sites of antibodies but lack the large crystallizable fragment of full-sized antibodies. Single chain nanobodies are entirely capable of binding to their antigen targets but are <10% of the mass of a human antibody (10-15Kda vs. 175Kda). The small molecular weight of these nanobodies makes them excellent candidates to accelerate structural biology approaches at PNNL including Native mass spectrometry and Cryo-EM. In native mass spectrometry experiments- these nanobodies hold promise to stabilize complexes during electrospray ionization and mass analysis without introducing some of the problems associated with employing full-sized antibodies (such as aggregation and precipitation). We have developed a platform to generate synthetic nanobodies at PNNL. We request instrument time to test two of these nanobodies on the Synapt G2 instrument or the UHMR orbital trap. Our objective is to determine if the nanobodies display improved electrospray and mass characterization against a full-sized monoclonal antibody. If the nanobodies display improved electrospray and mass resolution (compared to larger antibodies), then future EMSL users might employ similar nanobody platforms to stabilize multi-protein complexes for native mass spectrometry experiments.
Project Details
Start Date
N/A
End Date
N/A
Status
Accepted
Released Data Link
Team
Principal Investigator