Depletion of Albumin From Human Serum by Monosize Affinity Beads

Year 2013, Volume: 41 Issue: 3, 279 - 288, 01.09.2013

Evrim Banu Altıntaş

Abstract

The hydrophobic affinity ligand L-tryptophan immobilized magnetic poly glycidyl methacrylate [m-poly GMA ] beads in monosize form 1.6μm in diameter were used for the affinity depletion of human serum albumin [HSA]. The m-poly GMA beads were prepared by dispersion polymerization in the presence of Fe3O4 nano-powder. The epoxy groups of the m-poly GMA beads were converted into amino groups by using 1,6 diaminohexane i.e., spacer arm . L-tryptophan was then covalently immobilized on spacer arm attached m-poly GMA beads. Elemental analysis of immobilised L-tryptophan for nitrogen was estimated as 42.5 μmol/g polymer. Adsorption studies were performed under different conditions in a batch system i.e., medium pH, protein concentration and temperature . Maximum lysozyme adsorption amount of m-poly GMA and L-tryptophan immobilized m-poly GMA [m-poly GMA -L-tryptophan] beads were 1.78 and 172.9 mg/g, respectively. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated. It was also observed that after 5 adsorption–elution cycle, m-poly GMA -L-tryptophan beads can be used without significant loss in HSA adsorption capacity. The elution results demonstrated that the adsorption of HSA to the adsorbent was reversible.

Keywords

Magnetic Monosize Beads, Human Serum Albumin Depletion, Poly glycidyl methacrylate

Eş boyutlu manyetik affinite partiküller ile insan serumundan albumin uzaklaştırılması

Abstract

L -Triptofan immobilize edilmiş manyetik poli glisidil metakrilat 1.6 µm çapında partiküller [m-poly GMA ] insan serum albumin [HSA] serumdan uzaklaştırılmasında kullanılmıştır. m-poly GMA partiküller Fe3O4varlığında dispersiyon polimerizasyonu ile hazırlanmıştır. [m-poly GMA ] partiküllerin epoksi grupları 1,6 diaminohekzan kullanılarak amino gruplarına çevrilmiştir. İmmobilize edilmiş L-triptofan miktarı elemental analiz ile 42.5 µmol/g polimer olarak hesaplanmıştır. Kesikli deney sistemi kullanılarak çeşitli koşullarda örneğin ortam pH’ı, protein derişimi, sıcaklık adsorpsiyon çalışmaları yapılmıştır. Maksimum HSA adsorpsiyonu m-poly GMA ve L-triptofan immobilize m-poly GMA [m-poly GMA -L-triptofan partiküller için 1.78 ve 172.9 mg/g bulunmuştur. Yalancı birinci ve ikinci derece kinetic modellemeler uygulanmıştır. Ayrıca 5 defa adsorpsiyon-desorpsiyon çalışmasında sonra [m-poly GMA -L-triptofan partiküllerin HSA adsorpsiyon kapasinde düşüş olmadığı görülmüştür. Elüsyon çalışmaları HSA’nın adsorbente bağlanmasının geri dönüşümlü olduğunu göstermiştir

Keywords

Mikron Boyutlu Manyetik Partiküller, İnsan Serum Albumini, Poli glisidil metakrilat .

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