Česká společnost pro hmotnostní spektrometrii

(Česká konference hmotnostní spektrometrie 2018 - ThS-007)
In-situ enrichment and detection of biotinylated molecules by MALDI-compatible protein chips

Petra Darebná ^1,2, Petr Pompach ^1,3 *, Šárka Svobodová ², Michal Volný ^1,3, Petr Novák ^1,3

1. Institute of Microbiology, v.v.i., Czech Academy of Sciences, CZ-142 20 Prague, Czech Republic
2. Faculty of Science, Charles University in Prague, CZ-128 00 Prague, Czech Republic
3. AffiPro, s.r.o., CZ-250 63 Mratin, Czech Republic

Abstrakt

Immobilization of proteins is of fundamental interest in biochemistry, biosensor, and material science as well as bioanalytical chemistry. The presented technology allows preparation of MALDI compatible protein chips by ambient ion landing. The electrosprayed proteins immobilized on conductive surfaces allow a wide range of bioanalytical assays. The reaction takes place directly on the protein chip and is followed by in-situ analysis by MALDI mass spectrometry. The conductive surfaces were modified with biotin-binding proteins streptavidin, avidin and neutravidin by ambient ion landing utilizing a nanospray to produce protein ions and heated tube to dry them. The beam of charged species was deposited on the vertically mounted ITO slides that were kept at high voltage of the opposite polarity with respect to the spray. The ion landing of electrosprayed protein molecules was performed under atmospheric pressure by automated ion landing apparatus on dry metal and metal oxide surfaces. Compare to other materials, non-reactive surfaces suffer minimal nonspecific interactions with chemical species in the investigated sample and are thus an ideal substrate for selective protein chips. In this study, the performance of MALDI chips functionalized by avidin, neutravidin, and streptavidin for enrichment and detection of biotinylated proteins and peptides is demonstrated. The functionality of surfaces modified with biotin-binding molecules could help to drive the MALDI-TOF mass spectrometry toward its implementation into routine clinical practice.

* Korespondující autor: petrpompach@gmail.com

Poděkování:

This work was supported by Grant Agency of Charles University (1042217) and the Ministry of Education, Youth and Sports of CR within the LH15010, LD15089 and LQ1604 National Sustainability Program II (Project BIOCEV-FAR), the project ''BIOCEV'' (CZ.1.05/1.1.00/02.0109) and Biovendor a.s. Research Agreement.