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Масс-спектрометрическая идентификация и количественная оценка Ñ†ÐµÐ»ÐµÐ²Ñ‹Ñ Ð°Ð½Ñ‚Ð¸Ð³ÐµÐ½Ð¾Ð² в ÑÐ»Ð¾Ð¶Ð½Ñ‹Ñ Ð±ÐµÐ»ÐºÐ¾Ð²Ñ‹Ñ ÑÐ¼ÐµÑÑÑ

магистерская диссертация

Масс-спектрометрическая идентификация и количественная оценка Ñ†ÐµÐ»ÐµÐ²Ñ‹Ñ Ð°Ð½Ñ‚Ð¸Ð³ÐµÐ½Ð¾Ð² в ÑÐ»Ð¾Ð¶Ð½Ñ‹Ñ Ð±ÐµÐ»ÐºÐ¾Ð²Ñ‹Ñ ÑÐ¼ÐµÑÑÑ

Abstract

Целью данного исследования являлась разработка методики количественной оценки целевого антигена в сложной белковой смеси методом MALDI масс-спектрометрии с применением Ð¼ÐµÑ‡ÐµÐ½Ñ‹Ñ Ð¸Ð·Ð¾Ñ‚Ð¾Ð¿Ð¾Ð¼ кислорода 18O пептидов. Ð’ качестве сложной белковой смеси выступали лизированные клетки A549, заражённые вирусом гриппа А. Объектом исследования являлся вирусный неструктурный белок NS1, синтезирующийся в Ð·Ð°Ñ€Ð°Ð¶Ñ‘Ð½Ð½Ñ‹Ñ ÐºÐ»ÐµÑ‚ÐºÐ°Ñ Ð² процессе жизненного цикла вируса. Методику отработали на рекомбинантном белке NS1(1-124) и затем применили на Ð±ÐµÐ»ÐºÐ°Ñ Ð»Ð¸Ð·Ð¸Ñ€Ð¾Ð²Ð°Ð½Ð½Ñ‹Ñ ÐºÐ»ÐµÑ‚ÐºÐ°Ñ , предварительно Ñ€Ð°Ð·Ð´ÐµÐ»Ñ‘Ð½Ð½Ñ‹Ñ Ð¼ÐµÑ‚Ð¾Ð´Ð¾Ð¼ ЭФ в ПААГ.

The aim of this work was to develop a method of target antigen quantitative evaluation in a complex protein mixture by MALDI mass spectrometry using isotopomers, peptides that contain stable isotopes of oxygen. Cell lysates of A549 cells infected with influenza virus A were used as a complex protein mixture. The object of study was the NS1 protein. This protein is synthesized in the infected cells during the viral life cycle. The proposed procedure of quantitative evaluation was tested on purified protein NS1 and then applied to the lysed cells that had been previously separated by SDS-PAGE.

Keywords

вирусы гриппа, vaccine, Масс-спектрометрия, вакцины, influenza viruses, Белки

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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