A Novel Method For Multiplex Protein Biomarker Analysis Of Human Serum Using Quantitative MALDI Mass Spectrometry

Author(s):  
Aleksandr S. Taraskin ◽  
Konstantin K. Semenov ◽  
Alexey A. Lozhkov ◽  
Irina L. Baranovskaya ◽  
Aleksandr V. Protasov ◽  
...  
Biochimie ◽  
2021 ◽  
Vol 185 ◽  
pp. 87-95
Author(s):  
Aleksandr S. Taraskin ◽  
Konstantin K. Semenov ◽  
Aleksandr V. Protasov ◽  
Alexey A. Lozhkov ◽  
Aleksandr A. Tyulin ◽  
...  

Metabolites ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 253
Author(s):  
Kevin J. Zemaitis ◽  
Alexandra M. Izydorczak ◽  
Alexis C. Thompson ◽  
Troy D. Wood

The study of biological specimens by mass spectrometry imaging (MSI) has had a profound influence in the various forms of spatial-omics over the past two decades including applications for the identification of clinical biomarker analysis; the metabolic fingerprinting of disease states; treatment with therapeutics; and the profiling of lipids, peptides and proteins. No singular approach is able to globally map all biomolecular classes simultaneously. This led to the development of many complementary multimodal imaging approaches to solve analytical problems: fusing multiple ionization techniques, imaging microscopy or spectroscopy, or local extractions into robust multimodal imaging methods. However, each fusion typically requires the melding of analytical information from multiple commercial platforms, and the tandem utilization of multiple commercial or third-party software platforms—even in some cases requiring computer coding. Herein, we report the use of matrix-assisted laser desorption/ionization (MALDI) in tandem with desorption electrospray ionization (DESI) imaging in the positive ion mode on a singular commercial orthogonal dual-source Fourier transform ion cyclotron resonance (FT-ICR) instrument for the complementary detection of multiple analyte classes by MSI from tissue. The DESI source was 3D printed and the commercial Bruker Daltonics software suite was used to generate mass spectrometry images in tandem with the commercial MALDI source. This approach allows for the generation of multiple modes of mass spectrometry images without the need for third-party software and a customizable platform for ambient ionization imaging. Highlighted is the streamlined workflow needed to obtain phospholipid profiles, as well as increased depth of coverage of both annotated phospholipid, cardiolipin, and ganglioside species from rat brain with both high spatial and mass resolution.


2020 ◽  
Author(s):  
Aleksandr S. Taraskin ◽  
Konstantin K. Semenov ◽  
Aleksandr V. Protasov ◽  
Alexey A. Lozhkov ◽  
Alexandr A. Tyulin ◽  
...  

ABSTRACTOne of the main functions of alpha-2-macroglobulin (A2M) in human blood serum is the binding of all classes of protease. It is known that trypsin, after such interaction, possesses modified proteolytic activity. Trypsin first hydrolyzes two bonds in A2M’s ‘bait region’, and the peptide 705VGFYESDVMGR715 is released from A2M. In this work, specifics of the A2M-trypsin interaction were used to determine A2M concentration directly in human blood serum using MALDI mass-spectrometry. Following exogenous addition of trypsin to human blood serum in vitro, the concentration of the VGFYESDVMGR peptide was measured, using its isotopically-labeled analogue (18O), and A2M concentration was calculated. The optimized mass spectrometric approach was verified using a standard method for A2M concentration determination (ELISA) and the relevant statistical analysis methods. It was also shown that trypsin’s modified proteolytic activity in the presence of serum A2M can be used to analyze other serum proteins, including potential biomarkers of pathological processes. Thus, this work describes a promising approach to serum biomarker analysis that can be technically extended in several useful directions.


2018 ◽  
Author(s):  
Shazia Khan ◽  
Diego F Cobice ◽  
Dawn EW Livingstone ◽  
C Logan Mackay ◽  
Scott P Webster ◽  
...  

2020 ◽  
Vol 75 (14) ◽  
pp. 1749-1757
Author(s):  
E. A. Anikeenko ◽  
E. N. Rakhmatullina ◽  
D. I. Falev ◽  
O. Yu. Khoroshev ◽  
N. V. Ul’yanovskii ◽  
...  

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