Human IgG Adsorption by Spherical Albumin Biosorbents in SBF Media

Year 2014, Volume: 42 Issue: 4, 509 - 515, 01.11.2014

Aydan Gülsu Sabiha Demirci Fatma Ayhan Hakan Ayhan

Abstract

The objective of this study was to investigate the removal efficiency of immunoglobulin G IgG by albumin bi- osorbents in batch wise in four different simulated body fluid SBF solutions at room temperature. Albumin biosorbents were prepared as microsphere form by emulsion polymerization method. 0.1 g/ml albumin concentration, 1000 rpm stirring rate, 1% glutaraldehyde GA concentration 30 minutes crosslinking time were determined as optimal conditions in our previous study. The prepared biosorbents were then studied for IgG removal efficiency in four simulated body fluid SBF solutions [N Normal -SBF, G glucose -SBF, H hepes -SBF, T tris -SBF] at room temperature. IgG capturing capacity was obtained as 320 mg IgG/g with albümin biosor- bent 94,6% in N-SBF. It was observed that there was no leakage at washing with water. It may be concluded that prepared albumin biosorbents are sufficient in terms of efficiency of IgG removal. It was seem that this method is easy to perform and is useful as a first step in the detection of diagnostic markers in body fluids by applying proteomics technologies. We believe that albumin biosorbents offered the promising approach with good removal specificity and efficiency of IgG.

Keywords

Biosorbent, Albumin, IgG, SBF

Küresel Albumin Biyosorbentlerle SBF ortamlarında İnsan IgG Adsorpsiyonu

Abstract

B u çalışmanın amacı albümin biyosorbentlerin, kesikli sistemde dört farklı yapay vücut sıvısı ortamında İnsan immunglobulin G HIgG uzaklaştırma kapasitesinin araştırılmasıdır. Albumin biyosorbentler, emülsiyon polimerizasyon yöntemi ile mikroküre formunda hazırlandı. Denysel parametreler 0,1 g/ml albumin derişimi, 1000 rpm karıştırma hızı, %1 glutaraldehid derişimi ve 30 dakika çapraz bağlanma süresi optimum şartlar olarak daha önceki çalışmamızda belirlenmiştir. Daha sonra, hazırlanan biyosorbentlerle, oda sıcaklığında dört faklı yapay vücut sıvısı SBF çözeltisi [N Normal -SBF, G glukoz -SBF, H hepes -SBF, T tris -SBF] ortamında, HIgG uzaklaştırma çalışmaları yapılmıştır. Albumin biyosorbentlerle N-SBF ortamında HIgG yakalama kapasitesinin 320 mg HIgG/g %94,6 olduğu tespit edilmiştir. Saf su ile yıkama işlemlerinde kaçak olmadığı gözlemiştir. Hazırlanan albumin biyosorbentlerin HIgG uzaklaştırma verimliliği açısından yeterli olduğu sonucuna varılmıştır. Bu yöntemin, proteomik teknolojileri uygulayarak vücut sıvılarında teşhis belirteçlerinin saptanmasında, ilk adım olarak gerçekleştirilmesi, kolay ve yararlı olduğu görülmektedir. Albumin biyosorbentler, iyi bir uzaklaştırma seçiciliği ve etkinliği ile umut verici yaklaşım sunmaktadır

Keywords

Biosorbent, Albumin, IgG, SBF

References

• 1. C. Li, K.H. Lee, Affinity depletion of albumin from human cerebrospinal fluid using Cibacron-blue-3G-Aderivatized photopatterned copolymer in a microfluidic device. Anal. Biochem., 333 (2004) 381.
• 2. A. Kocourek, P. Eyckerman, B. Thome-Krome, The combined removal of albumin and immunoglobulins from human serum. Bio. Tech. Int., 17 (2005) 24.
• 3. J. Bailey, K. Zhang, N. Zolotarjova, G. Nicol, C. Szafranski, Removing high abundance proteins from serum. Genet. Eng. News, 2003, 23.
• 4. Affinity Chromatography Principles and Methods GE Healthcare p7-9 18-1022-29. Edition AD.
• 5. T. Kokubo, Bioactive glass ceramics: properties and applications. Biomaterials, 12 (1991) 155.
• 6. T. Kokubo, S. Ito, Z.T. Huang, T. Hayashi, S. Sakka, Ca, P-rich layer formed on high-strength bioactive glassceramic A-W. J. Biomed. Mater. Res., 24 (1990) 331.
• 7. S.B. Cho, K. Nakanishi, N.T. KokuboSoga, C. Ohtsuki, T. Nakamura, Apatite formation on silica gel in simulated body fluid: its dependence on structures of silica gels prepared in different media. J. Biomed. Mater. Res., 33 (1996) 145.
• 8. http://www.sigmaaldrich.com/Area_of_Interest/Life_ Science/Cell_Culture/Product_Lines/Classic_Media_ Salts.html.
• 9. Y. Song, D. Shan, R. Chen, F. Zhang, E.H. Han., Biodegradable behaviors of AZ31 magnesium alloy in simulated body fluid Mat Sci Eng C., 29 (2009) 1039.
• 10. Yang L, Zang E., Biocorrosion behavior of magnesium alloy in different simulated fluids for biomedical application, Mat Sci Eng C., 29 (2009) 1691.
• 11. A. Gülsu, Biyopolimer bazlı biyosorbentlerin sentezi, karakterizasyonu ve biyoafinite uygulamaları, Doktora Tezi, Muğla Üniversitesi Fen Bilimleri Enstitüsü, 2011.
• 12. S.T. Mathew, S.G. Devi, K.V. Sandhya, Formulation and Evaluation of Ketorolac Tromethamine-loaded Albumin Microspheres for Potential Intramuscular Administration, AAPS PharmSciTech., 8 (2007) 14.
• 13. E.B. Altıntaş, A. Denizli, Efficient Removal of Albumin from Human Serum by Monosize Dye Affinity Beads, J. Chromatogr. B, 832 (2006) 216.
• 14. D. Sitnikov, D. Chan, E. Thibaudeau, M. Pinard, J.M. Hunter, Protein depletion from blood plasma using a volatile buffer J. Chromatogr. B., 832 (2006) 41.
• 15. E.B. Altıntaş, N. Tüzmen, L. Uzun, A. Denizli, Immobilized Metal Affinity Adsorption for Monosize Beads, Ind. Eng. Chem. Res., 4 (2007) 7802.
• 16. N. Bereli, G. Şener, E.B. Altıntaş, H. Yavuz, A. Denizli, Poly(glycidyl methacrylate) beads embedded cryogels for pseudo-specific affinity depletion of albumin and immunoglobulin G, Mat. Sci, Engi C, 30 (2010) 323.
• 17. M.D. Seferovic, V. Krughkov, D. Pinto, V.K.M. Han, M.B. Gupta, Quantitative 2-D gel electrophoresisbased expression proteomics of albumin and IgG immunodepleted plasma, J. Chromatogr. B, 865 (2008) 147.
• 18. M. Karataş, S. Akgöl, H. Yavuz, R. Say, A. Denizli, Immunoglobulin G depletion from human serum with metal-chelated beads under magnetic field, Int. J. Biol. Macromol., 40 (2007) 254.
• 19. L. Uzun, C. Armutcu, O. Bicen, A. Ersoz, R. Say, A. Denizli, Simultaneous depletion of immunoglobulin G and albumin from human plasma using novel monolithic cryogel columns, Colloid Surface B., 112 (2013) 1.
• 20. L. Urbas, P. Brne, B. Gabor, M. Barut, M. Strlic, T.C. Petric, A. Strancar, Depletion of high-abundance proteins from human plasma using a combination of an affinity and pseudo-affinity column, J. Chromatogr. A, 1216 (2009) 2689.
• 21. G.W. Horgan Sample size and replication in 2D gel electrophoresis studies, J. Proteome Res., 6 (2007) 2884.
• 22. M.P. Molloy, E.E. Brzezinski, J.Q. Hang, M.T. McDowell, R.A. VanBogelen Overcoming technical variation and biological variation in quantitative proteomics, Proteomics, 3 (2003) 1912.