Metal chelate for protein adsorption studies pHEMA-GMA column filling materials including groups preparation

Year 2024, Volume: 13 Issue: 2, 522 - 527, 15.04.2024

Bahar Yılmaz Altınok Mukaddes Keskinateş Mevlüt Bayrakcı

https://doi.org/10.28948/ngumuh.1382364

Abstract

In this study, new poly(HEMA-co-GMA) or pHEMA-GMA support solid structures were prepared to help protein purification. The surface of the prepared pHEMA-GMA building materials was modified by binding different functional groups such as IDA and NTA. Nickel (Ni II) metal ion was passed through the modified cryogel structures. The monolithic cryogel structure chelated with Ni metal was characterized by SEM and FT-IR. GFP fluorescent protein binding studies of metal chelated cryogel structures were performed using UV-vis and fluorescence spectrophotometry. These cryogels were determined to bind 45% and 75% protein by UV-vis and fluorescence spectrophotometry. And these results were supported by visualization with the SDS PAGE method.

Keywords

p(HEMA-GMA), UV-vis and fluorescence spectrophotometer, SDS page, Nickel, IDA, NTA

Project Number

06-M-21

Protein adsorpsiyon çalışmaları için metal şelat grupları içeren pHEMA-GMA kolon dolgu malzemelerinin hazırlanması

Abstract

Bu çalışmada, protein saflaştırılmasına yardımcı olacak yeni poly(HEMA-co-GMA) veya pHEMA-GMA destek katı yapıları hazırlandı. Hazırlanan pHEMA-GMA yapı malzemelerin yüzeyi IDA ve NTA gibi farklı fonksiyonel grupların bağlanması ile modifiye edildi. Modifiye edilen kriyojel yapılarından nikel (Ni II) metal iyonu geçirildi. Ni metali ile şelatlanan monolitik kriyojel yapısı SEM ve FT-IR ile karakterize edildi. Metal şelatlı kriyojel yapıların GFP floresan protein bağlama çalışmaları UV-vis ve floresan spektrofotometresi ile yapıldı. Bu kriyojellerin UV-vis ve floresan spektrofotometresi ile %45 ve %75 oranında protein bağladıkları tespit edildi. Ve bu sonuçlar SDS PAGE yöntemi ile görselleştirilerek desteklendi.

Keywords

p(HEMA-GMA), UV-vis ve floresans spektrofotometresi, SDS page, Nikel, IDA, NTA

Ethical Statement

Yazarlar çıkar çatışması olmadığını beyan etmektedir.

Supporting Institution

Karamanoğlu Mehmetbey Üniversitesi, Bilimsel Araştırma Projeleri Komisyonu

Project Number

06-M-21

Thanks

06-M-21 numaralı projenin gerçekleştirilmesi için destek veren Karamanoğlu Mehmetbey Üniversitesi, Bilimsel Araştırma Projeleri Komisyonu’na teşekkür ederiz.

References

• H. Alkan, N. Bereli, Z. Baysal, A. Denizli, Antibody purification with protein A attached supermacroporous poly (hydroxyethyl methacrylate) cryogel. Biochemical Engineering Journal, 45(3), 201-208, 2009. https://doi.org/10.1016/j.bej.2009.03.013
• A. Baimenov, D. A. Berillo, S. G. Poulopoulos, V. J. Inglezakis, A review of cryogels synthesis, characterization and applications on the removal of heavy metals from aqueous solutions. Advances in colloid and interface science, 276, 102088, 2020. https://doi.org/10.1016/j.cis.2019.102088
• M. Bakhshpour, A. Derazshamshir, N. Bereli, A. Elkak, A. Denizli. [PHEMA/PEI]–Cu (II) based immobilized metal affinity chromatography cryogels: application on the separation of IgG from human plasma. Materials Science and Engineering: C, 61, 824-831, 2016. https://doi.org/10.1016/j.msec.2016.01.005
• M. Bayrakcı, B. Yılmaz, Biyoaktif giemsa boyama ajanı ile suda çözünen sulfonato kaliksaren moleküllerinin etkileşiminin spektrofotometrik olarak incelenmesi. Journal of the Institute of Science and Technology, 9(2), 985-992, 2019. https://doi.org/10.21597/jist.439066
• N. S. Bibi, N. K. Singh, R. N. Dsouza, M. Aasim, M. Fernández-Lahore, Synthesis and performance of megaporous immobilized metal-ion affinity cryogels for recombinant protein capture and purification. Journal of Chromatography A, 1272, 145– 149, 2013. https://doi.org/10.1016/j.chroma.2012.11.036
• B. M. A. Carvalho, L. H. M. Da Silva, L. M. Carvalho, A. M. Soares, L. A. Minim, S. L. Da Silva, Microcalorimetric study of the adsorption of lactoferrin insupermacroporous continuous cryogel with immobilized Cu2+ ions. Journal of Chromatography A, 1312, 1– 9, 2013. https://doi.org/10.1016/ j.chroma.2013.08.042
• M. B. Dainiak, F. M. Plieva, I. Y. Galaev, R. H. Kaul, B. Mattiasson, Cell Chromatography: Separation of Different Microbial Cells Using IMAC Supermacroporous Monolithic Columns. Biotechnology Progress, 21, 644-649, 2005. https://doi.org/10.1021/bp049615g
• N. Eczacioglu, B. Yilmaz, Y. Ulusu, M. Bayrakci, Recovery and reusability of apounag fluorescence protein from the unconjugated bilirubin complex structure. Journal of Fluorescence, 30(3), 497-503, 2020. https://doi.org/10.1007/s10895-020-02519-w
• K. Erol, B. K. Cebeci, K. Köse, D. A. Köse, Effect of immobilization on the activity of catalase carried by poly (HEMA-GMA) cryogels. International journal of biological macromolecules, 123, 738-743, 2019. https://doi.org/10.1016/j.ijbiomac.2018.11.121
• I. Guven, O. Gezici, M. Bayrakci, M. Morbidelli, Calixarene-immobilized monolithic cryogels for preparative protein chromatography. Journal of Chromatography A, 1558, 59–68, 2018. https://doi.org/10.1016/j.chroma.2018.05.026
• A. Hanora, F. Bernaudat, F. M. Plieva, M. B. Dainiak, L. Bülow, I. Y. Galaev, B. Mattiasson, Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format. Journal of Chromatography A, 1087, 38–44, 2005. https://doi.org/10.1016/j.chroma.2005.04.029
• S. A. A. Noma, Ö. Acet, A. Ulu, B. Önal, M. Odabaşı, B. Ateş, l-asparaginase immobilized p (HEMA-GMA) cryogels: A recent study for biochemical, thermodynamic and kinetic parameters. Polymer Testing, 93, 106980, 2021. https://doi.org/10.1016/j.polymertesting.2020.106980
• J. Porath, J. Carlsson, I. Olsson, G. Belfrage, Metal chelate affinity chromatography, a new approach to protein fractionation. Nature, 258, 598- 599, 1975.
• J. Sarkar, A. Kumar, Immobilized metal affinity cryogel-based high-throughput platform for screening bioprocess and chromatographic parameters of His6-GTPase. Analytical and Bioanalytical Chemistry, 409, 2951–2965, 2017. https://doi.org/10.1007/s00216-017-0242-9
• B. Wan, J. Li, F. Ma, N. Yu, W. Zhang, L. Jiang, H. Wei, Preparation and properties of cryogel based on poly (2-hydroxyethyl methacrylate-co-glycidyl methacrylate). Langmuir, 35(9), 3284-3294, 2019.: https://doi.org/10.1021/acs.langmuir.8b04021
• B. Yılmaz, Spektroskopik Teknikler Kullanılarak Furosemid ile DNA Etkileşimlerinin İncelenmesi. Bitlis Eren Üniversitesi Fen Bilimleri Dergisi, 10(2), 325-331, 2021. doi: https://doi.org/10.17798/bitlisfen.856526
• B. Yilmaz, Release of nifedipine, furosemide, and niclosamide drugs from the biocompatible poly (HEMA) hydrogel structures. Turkish Journal of Chemistry, 46(5), 1710-1722, 2022. https://doi.org/ 10.55730/1300-0527.3474
• Y. Şimşek, Sulu çözeltiden bakır (II) adsorpsiyon sürecinin optimizasyonunda yüzey yanıt metodolojisinin uygulanması. Academic Platform-Journal of Engineering and Science, 6(3), 182-191, 2018. https://doi.org/10.21541/apjes.393848
• M. Bayrakcı, B. Yılmaz, DNA ile Bağlanabilen Suda Çözünür Sülfonato Kaliks [8] Aren Sentezi ve Antimikrobiyal Aktivitesi. Niğde Ömer Halisdemir Üniversitesi Mühendislik Bilimleri Dergisi, 8(1), 615-620, 2019. https://doi.org/10.28948/ngumuh.517209
• M. Keskinates, B. Yilmaz, Y. Ulusu, M. Bayrakci, Electrospinning of novel calixarene-functionalized PAN and PMMA nanofibers: Comparison of fluorescent protein adsorption performance. Materials Chemistry and Physics, 205, 522-529, 2018. https://doi.org/10.1016/j.matchemphys.2017.11.055