Bruker's Ultraflex III Maldi-Tof/Tof instrument with its proprietary 200Hz smartbeam laser technology and the novel Maldi-TDS are unique in the market place for top-down recombinant protein sequencing
At Pittcon 2008, Bruker Daltonics introduced its powerful new and unique Maldi-TDS Tof/Tof based tools for biologics drug development and quality control in the pharmaceutical and biotechnology industries.
The Maldi-TDS (Maldi top-down solution) is available on Bruker's top-of-the line Ultraflex III Tof/Tof system for the characterisation of recombinant proteins with regard to sequence, C- and N-terminal assignments, Pegylation sites and other modifications.
Recently, the Maldi-TDS approach has been used to characterize two batches of recombinant proteins provided by the ABRF Protein Research Group (PRG) with the Ultraflex III's reISD (reflector in-source decay) Maldi top-down sequencing capability for unmatched 100% sequence coverage for all of the three different truncation variants to be determined.
The top-down analysis on the Ultraflex III system comprised simultaneously the assignment of all the N/C-termini of the recombinant proteins.
These data have been submitted to this year's ABRF-PRG 2008 study.
The robust reISD sequencing approach utilizes ultra-fast fragmentations in the Maldi ion source on the nanosecond time scale, providing predominantly C- and Z/Y-type fragment ions.
The Maldi-TDS tools also include targeted sequence analysis of N-terminal and C-terminal protein sequences by Bruker's proprietary T-sequencing technology.
In T-sequencing, fragment ions generated from near terminal regions of undigested proteins are further analysed in the Ultraflex III Tof/Tof, providing MS/MS spectra of protein terminal sequences.
The Maldi-TDS solves a major bottleneck in the development and quality control of biologics, namely the straightforward and unequivocal structure assignment of recombinant proteins for use as protein drugs.
The Maldi-TDS can also be applied for the characterization protein termini, even in the presence of terminal modifications, as well as for the characterization of Pegylated peptide and protein drugs, including the determination of PEGylation sites.
Detlev Suckau, Bruker Daltonics head of proteomics R+D, commented: "The characterisation of both N- and C-termini of proteins is a challenging undertaking, in particular for proteins present in mixtures.
"Our recent Ultraflex III Tof/Tof results presented in the ABRF study of top-down protein sequencing in combination with LC-Maldi separation of protein truncation variants were very impressive".
"It is clear that the new Maldi-TDS is a great complement to common LC-Maldi-Tof/Tof or LC-ESI/MS/MS bottom-up procedures, and can provide breakthrough new capabilities for pharma/biotech applications."
