nonlinear physics

“Early diagnosis of proteinopathies using massively parallel nano-spectroscopy with single-molecule sensitivity. Advanced clinical diagnostics for the development of personalized treatments”

Path to Precision Medicine project number: PPM 04-0131-200019

April 15th 2021 – April 15th 2024.

About the project

The project “Early diagnosis of proteinopathies using massively parallel nano-spectroscopy with single-molecule sensitivity. Advanced clinical diagnostics for the development of personalized treatments” is funded by the Qatar National Research Fund under grant number PPM 04-0131-200019.
The all-encompassing goal of this research project is to develop specialized photonics instrumentation based on the massively parallel Fluorescence Correlation Spectroscopy (mpFCS), mathematical analysis tools, and dedicated software that will allow us to translate our innovative method for ultrasensitive detection of structured peptide/protein aggregates to the primary care and use it to diagnose devastating proteopathy diseases with the ultimate, single-molecule sensitivity.

Aims of the project
  • Design and development of a compact, table-top instrument for mpFCS that will allow medical professionals to identify by screening individuals with elevated levels of structured peptide/protein aggregates in the blood serum, who are at risk to develop/have developed a proteopathy disease,
  • Design and development of a compact, table-top instrument for dual color mpFCS that will allow medical professionals to specifically diagnose the proteopathy disease in individuals who are identified by screening to have elevated levels of structured protein aggregates,
  • New innovative software for full control of single and dual color table-top instruments with fast acquisition, analysis and representation of large data sets.
Research methods

The idea to translate Fluorescence Correlation Spectroscopy (FCS), a quantitative analytical method with the ultimate, single-molecule sensitivity at room temperature [1] to healthcare, for rapid and cost-effective identification of individuals with elevated levels of structured peptide/protein aggregates in the blood serum (or other biological fluids), stems from our recent innovative work [2-4]. As a first step, we have shown that FCS in combination with Thioflavin T (ThT, figure A) is uniquely applicable for the detection of ThT-responsive structured aggregates in vitro [2] and in biological fluids (figure B) [3]. As a second step, we have successfully extended FCS to imaging and developed instrumentation for functional Fluorescence Microscopy Imaging (fFMI) by deploying simultaneous multi-point FCS [4] (figure C). Here, a Diffractive Optical Element (DOE) is used to generate an illumination matrix of 64×32, 32×32, or 16×16 spots that perfectly matches a 2D single-photon avalanche diode (SPAD) array, to simultaneously collect the signal in 1024 observation volume elements. This allows us to map the concentration, diffusion and velocity map of molecules of interest.

References
  1. Krmpot AJ, Nikolić SN, Oasa S, Papadopoulos DK, Vitali M, Oura M, Mikuni S, Thyberg P, Tisa S, Kinjo M, Nilsson L, Terenius L, Rigler R, Vukojević V.:
    “Functional Fluorescence Microscopy Imaging (fFMI). Quantitative Scanning-Free Confocal Fluorescence Microscopy for the Characterization of Fast Dynamic Processes in Live Cells”,
    Analytical Chemistry 91, 11129-11137 (2019).
  2. Tiiman A, Jelić V, Jarvet J, Järemo P, Bogdanović N, Rigler R, Terenius L, Gräslund A, Vukojević V.:
    “Amyloidogenic nanoplaques in blood serum of patients with Alzheimer’s disease revealed by time-resolved Thioflavin T fluorescence intensity fluctuation analysis”,
    J. Alzheimer’s Dis. 68, 571-582 (2019).
  3. Tiiman A, Jarvet J, Gräslund A, Vukojević V.:
    “Heterogeneity and intermediates turnover during amyloid-β (Aβ) peptide aggregation studied by Fluorescence Correlation Spectroscopy”,
    Biochemistry 54, 7203-7211 (2015).
  4. Bonito-Oliva A, Schedin-Weiss S, Younesi SS, Tiiman A, Adura C, Paknejad N, Brendel M, Romin Y, Parchem RJ, Graff C, Vukojević V, Tjernberg LO, Terenius L, Winblad B, Sakmar TP, Graham WV.:
    “Conformation-specific antibodies against multiple amyloid protofibril species from a single amyloid immunogen”,
    J Cell Mol Med. 23, 2103-2114 (2019).
Published papers with acknowledgement to the PPM project
  1. Sho Oasa, Aleksandar J. Krmpot, Stanko N. Nikolić, Andrew H. A. Clayton, Igor F. Tsigelny, Jean-Pierre Changeux, Lars Terenius, Rudolf Rigler, and Vladana Vukojević:
    “Functional Fluorescence Microscopy Imaging (fFMI). Quantitative Scanning-Free Confocal Fluorescence Microscopy for the Characterization of Fast Dynamic Processes in Live Cells”,
    Analytical Chemistry 91: 11129-11137 (2019).
Project team
texas dr belic
Dr. Milivoj R. Belić (Lead PI)

Office Phone: +974.4423.0124
Email: milivoj.belic@qatar.tamu.edu
Address: Texas A&M University at Qatar
Education City, Doha, Qatar

Vladana_image_1
Dr. Vladana Vukojević (PI)

Office Phone: +46-(0)8-517 717 22
Email: vladana.vukojevic@ki.se
Department of Clinical Neuroscience
Karolinska Institutet
Stockholm, Sweden

AKrmpot
Dr. Aleksandar J. Krmpot

Postdoctoral Research Associate
Email: aleksandar.krmpot@qatar.tamu.edu
Address: Texas A&M University at Qatar
Education City, Doha, Qatar

StankoNikolic
Dr. Stanko N. Nikolić

Postdoctoral Research Associate
Email: stanko.nikolic@qatar.tamu.edu
Address: Texas A&M University at Qatar
Education City, Doha, Qatar