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New Tricks to Sequence a Protein - Top-Down Mass Spectrometry and Biopharmaceutical Characterization

Nov. 09, 2011
Comparison of protein sequencing methods
Comparison of protein sequencing methods more
Comparison of protein sequencing methods Fig. 1: Schematics of MALDI Top-Down Protein Sequencing. The protein chain is fragmented by ... Fig. 2: MALDI Top-Down Sequence analysis of an antibody heavy chain. The N-terminal 77mer sequence ... Fig. 3: Analysis of recombinant Tau and its side products X and Y as they were detected in a ... Fig. 4: Sequence assignment of the N-terminal 65 residues the camelid nanobody R2P9 as compiled ... 
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Proteins become increasingly important in drug development, prompting for their detailed analysis and side product characterization in development and QC. Mass spectrometric Top-Down sequencing (TDS) addresses the key requirement to assign N- and C-terminal sequences in biologics and biosimilars. The principles of the new technology are described and related to the classical Edman-sequencing approach.

Therapeutic Protein Characterization
Numerous drugs used today are proteins (e.g. insulin or herceptin) and many more are in the development pipeline of the biopharmaceutical industry (1). They have to be characterized in terms of function, purity and structure with the same stringency as it has become standard for small molecular drugs (e.g., penicillin or aspirin) although this is much more difficult. This requires new technologies that help maintain drug safety even if the drug is, e.g., a large 150 kDa monoclonal therapeutic antibody, which is ­N-terminally pyroglutamylated, heterogeneously glycosylated and the C-terminal lysine residue at the heavy chain is removed.

Protein N-termini have been very successfully assigned and sequenced for the last 50 years by Edman sequencing (2). Protein C-termini, however, have never been amenable to routine chemical sequencing and were simply ignored, therefore. For protein drugs this is a critical issue and the new Top-Down protein sequencing capabilities that mass spectrometry provides will be very useful in this regard and are currently widely adopted in the biopharmaceutical industry.

Read more on Top-Down Mass Spectrometric Protein Sequencing within the pdf.

Teaser image: vege - Fotolia.com

 

Authors:
Anja Resemann
Arndt Asperger
Detlev Suckau

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Keywords : Anja Resemann Arndt Asperger Biopharmaceuts Biosimilars Bruker Daltonik Detlev Suckau Drug Development Edman-sequencing Life Science MALDI Mass spectrometric Top-Down sequencing Mass Spectrometry Proteomics Screening Application Note TDS

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