PHEMA Based Cryogel For Lectin Purification From Soybean Flour

Abstract

In this study poly hydroxyethyl methacrylate PHEMA based cryogel column was prepared and its usability for lectin purification from soybean flour was investigated. PHEMA cryogel was characterized by swelling test and scanning electron microscope SEM . N-acetyl-D-galactosamine GalNAc was immobilized to PHEMA cryogel as a ligand by cyanogen bromide activation. Soybean flour extract obtained from soybean seeds was used as source of lectin. Maximum lectin binding from soybean flour was 261 mg/g. Lectin elution from the N-acetyl-D-galactosamine immobilized PHEMA GalNAc-PHEMA cryogel was achieved by 0.5 M galactose. Purity of the lectin obtained from soybean flour was investigated by SDS-PAGE. Finally binding-elution cycle was performed for 5 times and no significant decrease was observed at column capacity.

Keywords

• PHEMA,cryogel,Lectins,Protein Purification,Soybean Flour

Soya Unundan Lektin Saflaştırılması için PHEMA Bazlı Kriyojel

Abstract

B u çalışmada poli hidroksietilmetakrilat PHEMA temelli kriyojel kolon hazırlanmış ve soya unundan lektin saflaştırılmasında kullanılabilirliği araştırılmıştır. PHEMA kriyojel taramalı elektron mikroskobu SEM ve şişme testi ile karakterize edilmiştir. PHEMA kriyojele ligand olarak N-asetil-D-galaktozamin siyanojen bromür CNBr aktivasyonu ile bağlanmıştır. Soya tohumlarından elde edilen soya unu özütü lektin kaynağı olarak kullanılmıştır. Soya unundan en fazla lektin bağlama miktarı 261 mg/g’dır. N-asetil-D-galaktozamin immobilize edilmiş PHEMA kriyojelden lektin elusyonu 0.5 M galaktoz ile gerçekleştirilmiştir. Soya unundan elde edilen lektinin saflığı SDS-PAGE ile araştırılmıştır. Son olarak, bağlanma ve elusyon döngüsü 5 kez tekrarlanmış ve kolon kapasitesinde önemli bir azalma gözlenmemiştir

Keywords

• PHEMA, Kriyojel, Lektinler, Protein Saflaştırma, Soya Unu.

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