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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

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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.

References

  1. Dong Z, Ahrens CC, Yu D, Ding Z, Lim HT, Li W (2017) Cell isolation and recovery using hollow glass microspheres coated with nanolayered films for applications in resource-limited settings. ACS Appl Mater Interfaces 9:15265–15273

    Article  CAS  PubMed  Google Scholar 

  2. Salimi K, Usta DD, Koçer İ, Çelik E, Tuncel A (2017) Highly selective magnetic affinity purification of histidine-tagged proteins by Ni2+ carrying monodisperse composite microspheres. RSC Adv 7:8718–8726

    Article  CAS  Google Scholar 

  3. Wang B, Shao Q, Fang Y, Wang J, Xi X, Chu Q, Dong G, Wei Y (2017) Fabrication of imidazolium-functionalized magnetic composite microspheres for selective recognition and separation of heme proteins. New J Chem 41:5651–5659

    Article  CAS  Google Scholar 

  4. Li D, Yi R, Tian J, Li J, Yu B, Qi J (2017) Rational synthesis of hierarchical magnetic mesoporous silica microspheres with tunable mesochannels for enhanced enzyme immobilization. Chem Commun 53:8902–8905

    Article  CAS  Google Scholar 

  5. Campos E, Branquinho J, Carreira AS, Carvalho A, Coimbra P, Ferreira P, Gil MH (2013) Designing polymeric microparticles for biomedical and industrial applications. Eur Polym J 49:2005–2021

    Article  CAS  Google Scholar 

  6. Wang C, Podgórski M, Bowman CN (2014) Monodisperse functional microspheres from step-growth “click” polymerizations: Preparation, functionalization and implementation. Mater Horizons 1:535–539

    Article  CAS  Google Scholar 

  7. Zhao Z-B, Tai L, Zhang D-M, Jiang Y (2017) Facile fabrication of siloxane@poly(methylacrylic acid) core-shell microparticles with different functional groups. J Nanopart Res 19:73

    Article  CAS  Google Scholar 

  8. Sathe TR, Agrawal A, Nie S (2006) Mesoporous silica beads embedded with semiconductor quantum dots and iron oxide nanocrystals: Dual-function microcarriers for optical encoding and magnetic separation. Anal Chem 78:5627–5632

    Article  CAS  PubMed  Google Scholar 

  9. Wang J, Yao J, Sun N, Deng C (2017) Facile synthesis of thiol-polyethylene glycol functionalized magnetic titania nanomaterials for highly efficient enrichment of N-linked glycopeptides. J Chromatogr A 1512:1–8

    Article  CAS  PubMed  Google Scholar 

  10. Zhang L, Yang Z, Zhu W, Ye Z, Yu Y, Xu Z, Ren J, Li P (2017) Dual-stimuli-responsive, polymer-microsphere-encapsulated CuS nanoparticles for magnetic resonance imaging guided synergistic chemo-photothermal therapy. ACS Biomater Sci Eng 3:1690–1701

    Article  CAS  Google Scholar 

  11. Omer-Mizrahi M, Margel S (2007) Synthesis and characterization of spherical and hemispherical polyepoxide micrometer-sized particles of narrow size distribution by a single-step swelling of uniform polystyrene template microspheres with glycidyl methacrylate. J Polym Sci Pol Chem 45:4612–4622

    Article  CAS  Google Scholar 

  12. Pei X, Zhai K, Liang X, Deng Y, Xu K, Tan Y, Yao X, Wang P (2018) Fabrication of shape-tunable macroparticles by seeded polymerization of styrene using non-cross-linked starch-based seed. J Colloid Interface Sci 512:600–608

    Article  CAS  PubMed  Google Scholar 

  13. Ling D, Hyeon T (2013) Chemical design of biocompatible iron oxide nanoparticles for medical applications. Small 9:1450–1466

    Article  CAS  PubMed  Google Scholar 

  14. Cornell RM, Schwertmann U (2000) The Iron Oxides: Structure, Properties, Reactions, Occurrences and Uses, 2nd edn. Wiley, Darmstadt

    Google Scholar 

  15. Laurent S, Forge D, Port M, Roch A, Robic C, Vander Elst L, Muller RN (2008) Magnetic iron oxide nanoparticles: Synthesis, stabilization, vectorization, physicochemical characterizations, and biological applications. Chem Rev 108:2064–2110

    Article  CAS  PubMed  Google Scholar 

  16. Myronovkij S, Negrych N, Nehrych T, Redowicz MJ, Souchelnytsky S, Stoika R, Kit Y (2016) Identification of a 48 kDa form of unconventional Myosin 1 C in blood serum of patients with autoimmune diseases. Biochem Biophys Rep 5:175–179

    PubMed  Google Scholar 

  17. Kubinová Š, Horák D, Syková E (2009) Cholesterol-modified superporous poly(2-hydroxyethyl methacrylate) scaffolds for tissue engineering. Biomaterials 30:4601–4609

    Article  CAS  PubMed  Google Scholar 

  18. Overberger CG, Oshaughnessy MT, Shalit H (1949) The preparation of some aliphatic azo nitriles and their decomposition in solution. J Am Chem Soc 71:2661–2666

    Article  Google Scholar 

  19. Porosimeter Pascal 140 and Pascal 440, Instruction manual (1996) p 8

  20. Rigby SP, Barwick D, Fletcher RS, Riley SN (2003) Interpreting mercury porosimetry data for catalyst supports using semi-empirical alternatives to the Washburn equation. Appl Catal A 238:303–318

    Article  CAS  Google Scholar 

  21. Groen JC, Peffer LAA, Pérez-Ramírez J (2003) Pore size determination in modified micro- and mesoporous materials. Pitfalls and limitations in gas adsorption data analysis. Microporous Mesoporous Mat 60:1–17

    Article  CAS  Google Scholar 

  22. Lowell S, Shields JE, Thomas MA, Thommes M (2004) Micropore analysis. characterization of porous solids and powders. In: Volume 16: Surface Area, Pore Size and Density, Particle Technology Series. Springer, New York, pp 129–156

  23. Horák D, Hlídková H, Kit Y, Antonyuk V, Myronovsky S, Stoika R (2017) Magnetic poly(2-hydroxyethyl methacrylate) microspheres for affinity purification of monospecific anti-p46 kDa/Myo1C antibodies for early diagnosis of multiple sclerosis patients. Biosci Rep 37, BSR20160526_1–BSR20160526_10

  24. Laemli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–385

    Article  Google Scholar 

  25. Rouquerol J, Avnir D, Fairbridge CW, Everett DH, Haynes JH, Pernicone N, Ramsay JDF, Sing KSW, Unger KK (1994) Recommendations for the characterization of porous solids. Pure Appl Chem 66:1739–1758

    Article  CAS  Google Scholar 

  26. Swern D (1970) Reactions of the oxirane group. J Am Oil Chem Soc 47:424–429

    Article  CAS  Google Scholar 

Download references

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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