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.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Atomic absorption spectroscopy
Brunauer-Emmett-Teller adsorption/desorption isotherm
- d :
Fourier transform infrared spectroscopy
- mag-PGMA-NH2 :
- PGMA-NH2 :
Poly(glycidyl methacrylate) microspheres modified with amino groups
- S BET :
Specific surface area
Scanning electron microscopy
Sodium dodecyl sulfate
SDS polyacrylamide gel electrophoresis
Transmission electron microscopy.
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
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
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
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
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
Wang C, Podgórski M, Bowman CN (2014) Monodisperse functional microspheres from step-growth “click” polymerizations: Preparation, functionalization and implementation. Mater Horizons 1:535–539
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
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
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
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
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
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
Ling D, Hyeon T (2013) Chemical design of biocompatible iron oxide nanoparticles for medical applications. Small 9:1450–1466
Cornell RM, Schwertmann U (2000) The Iron Oxides: Structure, Properties, Reactions, Occurrences and Uses, 2nd edn. Wiley, Darmstadt
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
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
Kubinová Š, Horák D, Syková E (2009) Cholesterol-modified superporous poly(2-hydroxyethyl methacrylate) scaffolds for tissue engineering. Biomaterials 30:4601–4609
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
Porosimeter Pascal 140 and Pascal 440, Instruction manual (1996) p 8
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
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
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
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
Laemli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–385
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
Swern D (1970) Reactions of the oxirane group. J Am Oil Chem Soc 47:424–429
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.
The authors declare no competing financial interest.
Electronic supplementary material
Rights and permissions
About this article
Cite this article
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
- Magnetic microspheres
- Affinity chromatography
- p46/Myo1C protein
- Immunoglobulin M
- Autoimmune disease marker
- Multiple sclerosis