Depletion of Albumin From Human Serum by Monosize Affinity Beads

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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