Adsorption of BSA on Metal Loaded Monosize p GMA Microspheres

Abstract

Immobilized metal ion affinity chromatography IMAC is an efficient method for the adsorption of histidine- having proteins so poly glycidyl methacrylate [p GMA ] monosize microspheres with an average diameter of 1.6 μm were produced by dispersion polymerization for adsorption of bovine serum albumin. Cu II ions were chelated on the poly glycidyl methacrylate -iminodiacetic acid [p GMA -IDA] microspheres to be used in bovine serum albumin adsorption studies in batch system. Characterizations of the microspheres were carried out by scanning electron microscopy SEM , elemental analysis, Fourier transform infrared FTIR spectroscopy. The maximum adsorption capacity of the p GMA -IDA-Cu II microspheres was found to be 278.6 mg/g polymer at pH 5.0. The elution studies were performed by 1.0 M NaCl solution. The obtained results showed that the metal chelated p GMA -IDA-Cu II microspheres can be evaluated as an efficicent adsorbent for albumin adsorption.

Keywords

• Immobilized metal ion affinity chromatography,Albumin,Proteomics,Poly glycidyl methacrylate

Metal Yüklenmiş Eşboyutlu p GMA mikrokürelere BSA Adsorpsiyonu

Abstract

İ mmobilize metal iyon afinite kromatografi İMAK , histidin taşıyan proteinlerin adsorpsiyonu için etkili bir yöntem olduğundan sığır serum albuminin uzaklaştırılması için dispersiyon polimerizasyon yöntemiyle, 1.6 µm çapında eş boyutlu poli glisidil metakrilat [p GMA ] mikroküreler üretilmiştir. Cu II iyonları, kesikli sistemde sığır serum albuminin adsorpsiyon çalışmalarında kullanılmak üzere p GMA -IDA mikroküreleri ile şelatlanmıştır. Mikrokürelerin karakterizasyonu taramalı elektron mikroskobu SEM , elementel analiz, Fourier dönüşüm kızılötesi FTIR spektroskopisi kullanılarak gerçekleştirilmiştir. p GMA -IDA-Cu II mikrokürelerinin maksimum adsorpsiyon kapasitesi pH 5.0’te 278.6 mg/g polimer bulunmuştur. Elüsyon çalışmaları 1.0 M NaCl çözeltisi ile gerçekleştirilmiştir. Elde edilen sonuçlar p GMA -IDA-Cu II mikrokürelerin albüminin adsorpsiyonu için etkin bir adsorban olarak kabul edilebileceğini göstermiştir

Keywords

• İmmobilize metal iyon afinite kromatografi,Albumin,Proteomik,Poli glisidil metakrilat

References

1. J. Metcoff, F.J. Stare, The Physiologic and Clinical Significance of Plasma Proteins and Protein Metabolites, New England Journal of Medicine. 236 (1947) 68-75.
2. N.L. Anderson, N.G. Anderson, The human plasma proteome: history, character, and diagnostic prospects, Mol Cell Proteomics. 1 (2002) 845-67.
3. H.H. Yang, K.H. Lu, Y.F. Lin, S.H. Tsai, S. Chakraborty, W.J. Zhai, D.F. Tai, Depletion of albumin and immunoglobulin G from human serum using epitope- imprinted polymers as artificial antibodies, Journal of Biomedical Materials Research Part A. 101A (2013) 1935-1942. magnetic microspheres for the recognition of albumin, Journal of Separation Science. 37 (2014) 2077-2086.
4. S. Kovac, X. Yang, F. Huang, D. Hixson, D. Josic, Proteomics as a tool for optimization of human plasma protein separation, Journal of Chromatography A. 1194 (2008) 38-47.
5. E.J. Cohn, L.E. Strong, W.L. Hughes, D.J. Mulford, J.N. Ashworth, M. Melin, H.L. Taylor, Preparation and Properties of Serum and Plasma Proteins. IV. A System for the Separation into Fractions of the Protein and Lipoprotein Components of Biological Tissues and Fluids, Journal of the American Chemical Society. 68 (1946) 459-475.
6. A. Denizli, Plasma fractionation: conventional and chromatographic methods for albumin purification, Hacettepe Journal of Biology and Chemistry. 39 (2011) 315-341.
7. R. Gutiérrez, E.M.M. Del Valle, M.A. Galán, Adsorption and mass transfer studies of Catalase in IMAC chromatography by dynamics methods, Process Biochemistry. 41 (2006) 142-151.
8. N. Bereli, G. Şener, E. B. Altıntaş, H. Yavuz, A. Denizli, Poly(glycidyl methacrylate) beads embedded 16. J. Porath, Immobilized metal ion affinity cryogels for pseudo-specific affinity depletion of albumin and immunoglobulin G, Materials Science and Engineering C. 30 (2010) 323–329.

9. O. González-Ortega, J. Porath, R. Guzmán, Adsorption of peptides and small proteins with control access polymer permeation to affinity binding sites. Part I: Polymer permeation-immobilized metal ion affinity chromatography separation adsorbents with polyethylene glycol and immobilized metal ions. Journal of Chromatography A. 1227 (2012)115-125.
10. J.T. Mooney, D.P. Fredericks, M.T.W. Hearn, Application of an IMAC cassette for the purification of N-terminally tagged proteins, Separation and Purification Technology. 120 (2013) 265-274.
11. R.R. Prasanna, M.A. Vijayalakshmi, Characterization of metal chelate methacrylate monolithic disk for purification of polyclonal and monoclonal immunoglobulin G, Journal of Chromatography A. 1217 (2010) 3660-7.
12. V. Karakoç, E. Yılmaz, D. Türkmen, N. Öztürk, S. Akgöl, A. Denizli, Selective separation of human serum albumin with copper(II) chelated poly(hydroxyethyl methacrylate) based nanoparticles, International Journal of Biological Macromolecules. 45 (2009) 188–193.
13. M.B. Ribeiro, M. Vijayalakshmi, D. Todorova-Balvay, S.M.A. Bueno, Effect of IDA and TREN chelating agents and buffer systems on the purification of human IgG with immobilized nickel affinity membranes, Journal of Chromatography B. 861 (2008) 64-73.
14. P. Jain, L. Sun, J. Dai, G.L. Baker, M.L. Bruening, High-Capacity Purification of His-tagged Proteins by Affinity Membranes Containing Functionalized Polymer Brushes, Biomacromolecules. 8 (2007) 3102-3107. chromatography, Protein Expression and Purification. 3 (1992) 263-281.
15. E.K.M. Ueda, P.W. Gout, L. Morganti, Current and prospective applications of metal ion–protein binding, Journal of Chromatography A. 988 (2003) 1-23.
16. V. Gaberc-Porekar, V.Menart, Potential for Using Histidine Tags in Purification of Proteins at Large Scale, Chemical Engineering & Technology. 28 (2005) 1306-1314.
17. J.S. Mandeville, H.A. Tajmir-Riahi, Complexes of Dendrimers with Bovine Serum Albumin, Biomacromolecules. 11 (2010) 465–472.
18. T.S. Anirudhan, S.R. Rejeena, A.R. Tharun, Preparation, characterization and adsorption behavior of tannin- modified poly(glycidylmethacrylate)-grafted zirconium oxide-densified cellulose for the selective separation of bovine serum albumin, Colloids and Surfaces B: Biointerfaces. 93 (2012) 49-58.
19. E.B. Altıntaş, A. Denizli, Monosize poly(glycidyl methacrylate) beads for dye-affinity purification of lysozyme, International Journal of Biological Macromolecules. 38 (2006) 99-106.
20. Y. Wang, G. Wang, Y. Xiao, Y. Yang, R.Tang, Yolk–Shell Nanostructured Fe3O4@NiSiO3 for Selective Affinity and Magnetic Separation of His-Tagged Proteins, ACS Applied Materials & Interfaces. 6 (2014) 19092-19099.
21. F.H. Arnold, Metal-affinity separations: a new dimension in protein processing, Biotechnology (N Y). 9 (1991) 151-6.