When genetic source material is unavailable, de novo sequencing by mass spectrometry is the direct path to sequence recovery.
CovalX published a case study with Green Mountain Antibodies demonstrating this workflow end to end: from purified antibody to functional recombinant, in 4 weeks.
Two reference antibodies were sequenced (trastuzumab, anti-Factor XII), expressed in Expi293 cells, and characterized by SEC, SDS-PAGE, ELISA, and BLI. Both showed monomer content above 96% and binding affinity consistent with the originals.
Key technical points:
· Multi-enzyme parallel digestion for maximum peptide redundancy
· nLC-Orbitrap MS/MS with EThCD for direct Ile/Leu distinction
· 100% sequence coverage on heavy and light chains
· PTMs reported, N-glycans included