Preparation of an affinity cryogel column for lysozyme purification

Abstract

Affinity cryogels were synthesized using tris(hydroxymethyl)aminomethane (Tris) as ligand for specific interactions with the lysozyme (LYZ). The cryogel was produced by cryocopolymerization at −12 °C. A central composite rotational design 2 2 was used to optimize the immobilization procedure of the Tris on the cryogel. A maximum adsorption for LYZ (149.5 mg/g) was achieved when 376 mg/mL of Tris and 3.06 mol/L of sodium borohydride were used during the Tris immobilization. Chromatographic separation of LYZ from chicken egg white was done with a purity of 92.13 %. Results showed that the affinity cryogel was a potential separation medium for LYZ purification.

Preparation of supermacroporous cryogels with improved mechanical strength for efficient purification of lysozyme from chicken egg white

“…The average value of the maximum adsorptive capacity was approximately 360 mg/g for the three Tris‐cryogels in this work, which was comparable to or greater than the values reported in the literature [42,44,55‐59]. As shown in Table 2, the Tris ligand exhibited the highest affinity for lysozyme compared with the other ligands.…”

“…Moreover, compared to the two electrophoretic bands of effluents each from the Tris‐pAAm and Tris‐pHEMA cryogels, the obtained effluent from the Tris‐pAAm‐co‐HEMA cryogel exhibited a single and clear electrophoretic band around 14.7 kD of lysozyme (Fig. 6B) [55]. This was attributed to the relatively small pores of the Tris‐pAAm and Tris‐pHEMA cryogels, which led to difficulty in impurity clearance.…”

Funcionalização De Criogéis Com Polietilenoimina Para a Adsorção Da Albumina Do Soro Bovino Por Cromatografia De Troca Iônica

“…A produção de criogel foi realizada de acordo com a metodologia proposta por Veríssimo et al (2017). Uma mistura contendo 1,185 g de acrilamida e 0,3175 g de Nmetileno-bis-acrilamida e 1 mL de alilglicidil éter foi solubilizada em 25 mL de água deionizada.…”

“…Entretanto, sua grande porosidade faz com que este apresente uma menor área superficial disponível para adsorção quando comparado a um leito fixo empacotado, surgindo a necessidade de modificações em sua estrutura para aumentar a eficiência da separação. Isto pode ser alcançado por meio da imobilização de ligantes na superfície do criogel, visando a captura seletiva do soluto-alvo (LOZINSKY et al, 2001;VERÍSSIMO et al, 2017).O presente trabalho teve como objetivo imobilizar o grupo ligante polietilenoimina (PEI) na superfície de criogéis supermacroporosos, visando a adsorção da albumina do soro bovino…”

Poli(Hidroksietilmetakrilat-Genişletilmiş Perlit) Kompoziti ile Yumurta Akından Lizozimin Tek Basamakta Saflaştırılması

“…There are wide range of methods that are used to isolate lysozyme. Some of these methods are: Aqueous biphasic system for lysozyme separation studies [4], affinity cryogel and composite cryogel for lysozyme purification [5][6][7]. In addition, dye-attached nanoparticles were used for lysozyme seperation [8].…”