Naslov
Data independent acquisition de novo sequencing technique

Autori
Dončević, Lucija ; Biba, Renata ; Cindrić, Mario

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
28HSKIKI : 28th Croatian Meeting of Chemists and Chemical Engineers and 6th Symposium Vladimir Prelog : Book of Abstracts / Rogošić, Marko - Zagreb : Hrvatsko društvo kemijskih inženjera i tehnologa (HDKI), 2023, 68-68

Skup
28HSKIKI - 28th Croatian Meeting of Chemists and Chemical Engineers and 6th Symposium Vladimir Prelog

Mjesto i datum
Rovinj, Hrvatska, 28.03.2023. - 31.03.2023

Vrsta sudjelovanja
Poster

Vrsta recenzije
Podatak o recenziji nije dostupan

Ključne riječi
de novo sequencing ; data dependent acquisition ; data independent acquisition ; chemically activated fragmentation

Sažetak
Peptide or protein de novo sequencing, method that can acquire peptide and protein sequence without the assistance of a protein database, represents one of the most challenging tasks in proteomics. Peptide or protein mass spectra can be produced using either data dependent or data independent acquisition (DDA or DIA). DDA is selective, more informative and sensitive in terms of a single ion, and thus represents a method of choice for de novo sequencing. Compared to DDA, DIA is faster, easier to use and optimize, and it dissociates and analyzes all present ions. However, it presents a couple of drawbacks. Firstly, the de novo sequencing algorithm is not always able to deliver a complete sequence or it has uncertainty in a portion of the derived sequence. Moreover, peptide or protein identification score results are significantly lower compared to DDA. To overcome named disadvantages, development of different approaches in de novo sequencing techniques is required. In this research, chemically activated fragmentation (CAF) coupled with charge reduction conditions was applied to enable directed peptide fragmentation of bovine serum albumin (BSA) tryptic peptides for the production of y- and b-ions in positive and negative mass spectrometer ion mode, respectively. Peptide ion spectra obtained by DIA analysis showed that 96.2% of the BSA protein sequence was acquired in comparison to 41.0% sequence coverage provided by the conventional DDA analysis. Implementation of this methodology could improve the quality of de novo sequencing dataset and provide unambiguous identification results.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

POVEZANOST RADA