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MASS SPECTROMETRY
AND PROTEOMICS
International Conference on
J u n e 2 5 - 2 7 , 2 0 1 8 | D u b l i n , I r e l a n d
Journal of Systems Biology & Proteome Research
|
Volume 2
Page 16
Note:
D
e-novo
, full-length sequencing of unknown proteins such as antibodies or
constituents of metaproteomes remains a challenging problem. Traditional
‘bottom-up’ proteomics approaches use proteolytic digestion, LC-MS/MS and
database searching to elucidate peptide identities and their parent proteins.
Protein sequences absent from the database cannot be identified, and even
if present in the database, complete sequence coverage is rarely achieved
even for the most abundant proteins in the sample. To this aim we have
developed Database Independent Protein Sequencing (DiPS), a novel method
for unambiguous, rapid, database independent, full-length protein sequencing.
The method is based on non-enzymatic, semi-random cleavage of the protein
by microwave assisted acid hydrolysis (MAAH), LC-MS/MS analysis, peptide
de novo sequencing, extraction of peptide tags, and their assembly into a
consensus sequence using a novel algorithm named Peptide Tag Assembler
(pTA). The method, which was recently published, now also allows for
differentiation between the isobaric leucine and isoleucine residues, and was
successfully applied to a variety of proteins and clinically relevant antibodies.
Biography
Alon Savidor completed his PhD at 2008 from the
University of Tennessee and The Oak Ridge Na-
tional Laboratory at Tennessee, USA. He complet-
ed his post-doctorate fellowship at the Tel Aviv
University, Israel, at 2013, and since then he is a
staff scientist at the Nancy and Stephen Grand
Israel National Center for Personalized Medicine,
Weizmann Institute of Science, Israel.
alon.savidor@weizmann.ac.ilDATABASE INDEPENDENT PROTEIN
SEQUENCING (DIPS) ENABLES
FULL-LENGTH DE-NOVO PROTEIN
AND ANTIBODY SEQUENCE
DETERMINATION
Alon Savidor
Weizmann Institute of Science, Israel
Alon Savidor, J Syst Biol Proteome Res 2018, Volume 2