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Monodisperse magnetic poly(glycidyl methacrylate) microspheres for isolation of autoantibodies with affinity for the 46 kDa form of unconventional Myo1C present in autoimmune patients

Microchimica Acta volume 185, Article number: 262 (2018) Cite this article

Abstract

Monodisperse nonmagnetic macroporous poly(glycidyl methacrylate) (PGMA) microspheres were synthesized by multistep swelling polymerization of glycidyl methacrylate, ethylene dimethacrylate and 2-[(methoxycarbonyl)methoxy]ethyl methacrylate (MCMEMA). This was followed (a) by ammonolysis to modify the microspheres with amino groups, and (b) by incorporation of iron oxide (γ-Fe2O3) into the pores to render the particles magnetic. The resulting porous and magnetic microspheres were characterized by scanning and transmission electron microscopy (SEM and TEM), atomic absorption and Fourier transform infrared spectroscopy (AAS and FTIR), elemental analysis, vibrating magnetometry, mercury porosimetry and Brunauer-Emmett-Teller adsorption/desorption isotherms. The microspheres are meso- and macroporous, typically 5 μm in diameter, contain 0.9 mM · g−1 of amino groups and 14 wt.% of iron according to elemental analysis and AAS, respectively. The particles were conjugated to p46/Myo1C protein, a potential biomarker of autoimmune diseases, to isolate specific autoantibodies in the blood of patients suffering from multiple sclerosis (MS). The p46/Myo1C loaded microspheres are shown to enable the preconcentration of minute quantities of specific immunoglobulins prior to their quantification via SDS-PAGE. The immunoglobulin M (IgM) with affinity to Myo1C was detected in MS patients.

Monodisperse magnetic poly(glycidyl methacrylate) microspheres were synthesized, conjugated with 46 kDa form of unconventional Myo1C protein (p46/Myo1C) via carbodiimide (DIC) chemistry, and specific autoantibodies isolated from blood of multiple sclerosis (MS) patients; immunoglobulin M (IgM) level increased in MS patients.

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Abbreviations

AAS:

Atomic absorption spectroscopy

BET:

Brunauer-Emmett-Teller adsorption/desorption isotherm

d :

Pore diameter

FTIR:

Fourier transform infrared spectroscopy

GMA:

Glycidyl methacrylate

MCMEMA:

2-[(Methoxycarbonyl)methoxy]ethyl methacrylate

mag-PGMA-NH2 :

Magnetic PGMA-NH2

MS:

Multiple sclerosis

PGMA:

Poly(glycidyl methacrylate)

PGMA-NH2 :

Poly(glycidyl methacrylate) microspheres modified with amino groups

S BET :

Specific surface area

SEM:

Scanning electron microscopy

SDS:

Sodium dodecyl sulfate

SDS-PAGE:

SDS polyacrylamide gel electrophoresis

TEM:

Transmission electron microscopy.

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Acknowledgements

Support of LQ1604 NPU II provided by MEYS and CZ.1.05/1.1.00/02.0109 BIOCEV provided by ERDF and MEYS, the Cedars-Sinai Medical Center’s International Research and Innovation Management Program and the RECOOP HST Association, as well as Volkswagen Foundation Trilateral Partnership Ukraine, Russian Federation and Germany, and the Target Complex Interdisciplinary Research Program of the NAS of Ukraine (No. 32-16) is acknowledged. The authors thank to Ms. Jiřina Hromádková for the micrographs of the particles.

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Authors and Affiliations

  1. Department of Polymer Particles, Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06, Prague 6, Czech Republic

    Beata A. Zasońska, Helena Hlídková & Daniel Horák

  2. Geomagnetic Department, Institute of Geophysics, Academy of Sciences of the Czech Republic, Boční II/1401, 141 31, Prague 4, Czech Republic

    Eduard Petrovský

  3. Department of Regulation of Cell Proliferation and Apoptosis, Institute of Cell Biology, NAS of Ukraine, Drahomanov Str. 14/16, Lviv, 79005, Ukraine

    Severyn Myronovskij, Volodymyr Antonyuk, Rostyslav Stoika & Yuriy Kit

  4. Department of Neurology, Danylo Halytsky Lviv National Medical University, Pekarska Str. 69, Lviv, 79010, Ukraine

    Tetyana Nehrych, Nazar Negrych & Mariya Shorobura

Authors
  1. Beata A. Zasońska
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  2. Helena Hlídková
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  3. Eduard Petrovský
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  4. Severyn Myronovskij
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  5. Tetyana Nehrych
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  6. Nazar Negrych
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  7. Mariya Shorobura
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  8. Volodymyr Antonyuk
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  9. Rostyslav Stoika
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  10. Yuriy Kit
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  11. Daniel Horák
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Correspondence to Daniel Horák.

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Zasońska, B.A., Hlídková, H., Petrovský, E. et al. Monodisperse magnetic poly(glycidyl methacrylate) microspheres for isolation of autoantibodies with affinity for the 46 kDa form of unconventional Myo1C present in autoimmune patients. Microchim Acta 185, 262 (2018). https://doi.org/10.1007/s00604-018-2807-5

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  • DOI: https://doi.org/10.1007/s00604-018-2807-5

Keywords

  • Magnetic microspheres
  • Functionalization
  • Affinity chromatography
  • p46/Myo1C protein
  • Autoantibody
  • Immunoglobulin M
  • Autoimmune disease marker
  • Multiple sclerosis