Affinity membrane chromatography: relationship of dye-ligand type to surface polarity and their effect on lysozyme separation and purification

Arıca, M. Yakup; Yılmaz, Meltem; Yalçın, Emine; Bayramoğlu, Gülay

doi:10.1016/j.jchromb.2004.03.021

Cited by 78 publications

(39 citation statements)

References 38 publications

(40 reference statements)

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“…9, it was observed that the binding capacity was increased from 154.3 to 309.2 mg/g as the temperature increased from 277 to 323 K. An increase in temperature caused an increase in binding capacity. Similar results were also observed in the adsorption of lysozyme by other dyed membranes (27,(39)(40)(41)(42)(43). The increase in binding capacity of the membrane was related to the conformational changes of the lysozyme molecule.…”

Section: Influence Of Temperaturesupporting

confidence: 80%

Dye Affinity Nanofiber Membrane for Adsorption of Lysozyme: Preparation and Performance Evaluation

Wang

Yang

Hsin

et al. 2017

Food Technol. Biotechnol.

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SUMMARYPolyacrylonitrile (PAN) nanofibrous membrane was prepared by an electrospinning technique. After heat treatment and alkaline hydrolysis, the weak ion exchange membrane was grafted with chitosan molecule and then covalently immobilized with a Cibacron Blue F3GA (CB). Fibre diameter, porosity and pore size of the membrane and immobilized dye density were characterized. Furthermore, the membrane was applied to evaluate the binding performance of lysozyme under various operating parameters (pH, chitosan mass per volume ratio, dye concentration, ionic strength and temperature) in batch mode. The experimental results were directly applied to purify lysozyme from chicken egg white by membrane chromatography. The results showed that the capture efficiency, recovery yield and purification factor were 90 and 87 %, and 47-fold, respectively, in a single step. The binding capacity remained consistent after five repeated cycles of adsorption-desorption operations. This work demonstrates that the dye-affinity nanofibrous membrane holds great potential for purification of lysozyme from real feedstock.

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Section: Influence Of Temperaturesupporting

confidence: 80%

Dye Affinity Nanofiber Membrane for Adsorption of Lysozyme: Preparation and Performance Evaluation

Wang

Yang

Hsin

et al. 2017

Food Technol. Biotechnol.

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“…To address this issue, we attempted to purify lysozyme using another method: ion exchange chromatography as was initially proposed by Strang (1984) and further modified by Luding et al (2011), Safarik et al (2007), Arica et al (2004) and Li & Chen (2002), just to name a few. The method is theoretically feasible to be used as a single step to purify EWL on the basis of isoelectric point (pI) of EWL is extremely higher (10.7) compared to that of other egg white proteins (< 6.5) (Anton et al, 2006;Machado et al, 2007;Luding et al, 2011).…”

Section: Purification Lysozymementioning

confidence: 99%

Purification of Egg White Lysozyme from Indonesian Kampung Chicken and Ducks

Wulandari

Fardiaz²,

Budiman³

et al. 2015

Med Pet

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Egg white lysozyme (EWL) has considerably a wide functional protein exhibiting antibacterial activity mainly against Gram-positive bacteria. The EWL is widely applied in food industry and is considerably safe. Despite its high potency, EWL of Indonesian poultry has never been studied and exploited. This study was aimed to purify EWL from two Indonesian poultry: kampung chicken and Cihateup duck, and compared to egg of commercial laying hens. The eggs in this study were obtained from field laboratory of Faculty of Animal Science, Bogor Agricultural University (IPB) and classified in AA quality based on the interior quality. First attempt to purify the EWL was performed by using ethanol precipitation yielding purified EWL which was still contaminated by other proteins, hence designated as partially purified EWL. Final concentrations of partially purified EWL of kampung chicken, commercial laying hens, and Cihateup duck were about 5800, 5400, and 5500 μg/mL, respectively. To confirm whether the use of ethanol in the purification affecting EWL antibacterial activities, the activities were examined against Staphylococcus aureus. It demonstrated that the partially purified EWL exhibited ability to inhibit S. aureus at 6 and 26 h suggesting that the method was feasible as it did not interfere EWL antibacterial activities. Yet, based on SDS-Page, purity was the issue in ethanol precipitation method. Further attempt using ion exchange chromatography at pH 10 successfully purified lysozyme as indicated by a single band corresponding to lysozyme size (~14 kD) free from bands of other proteins. Altogether, a single step of ion exchange chromatography is sufficient and promising to isolate EWL from Indonesian poultry for various industrial purposes.Key words: Indonesian poultry, lysozyme, egg, kampung chicken, Cihateup duck ABSTRAK Lisozim putih telur memiliki fungsi yang luas, salah satunya adalah memiliki aktivitas sebagai antibakteri terutama terhadap bakteri Gram positif. Lisozim dari putih telur �uga banyak diapli-. Lisozim dari putih telur �uga banyak diaplikasikan di industri pangan dan dinyatakan aman untuk pangan. Sampai saat ini lisozim dari putih telur yang berasal dari unggas lokal (Indonesia) belum dipela�ari dan dikembangkan. Tu�uan penelitian ini adalah melakukan purifikasi putih telur dari telur unggas lokal, yaitu ayam kampung dan itik Cihateup dibandingkan dengan telur ayam ras. Sampel telur yang didapatkan dari Laboratorium Lapang Fakultas Peternakan diklasifikasikan pada grade AA berdasarkan kualitas interior. Tahap pertama purifikasi lisozim putih telur dengan presipitasi etanol menghasilkan protein lain selain lisozim (purifikasi parsial). Konsentrasi lisozim putih telur hasil purifikasi parsial telur ayam kampung, ayam ras dan itik Cihateup masing-masing adalah 5800, 5400, dan 5500 ug/mL. Untuk melihat efek penggunaan etanol dalam proses purifikasi terhadap aktivitast antibakteri lisozim, kemampuan daya hambat lisozim terhadap Staphylococcus aureus telah dianalisis. Lisozim putih telur hasil pu...

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“…After the diluted solution was immersed in an ice bath and centrifuged at 10 000 rpm for 30 minutes, the supernatant solution was filtered and stored at 0 8C prior to use as a lysozyme source. [42] The effective separation of lysozyme from the CEW on the MIP3 monolith is shown in Figure 9. A protein peak was seen in Figure 9, which flowed later than other existing proteins on the MIP3 monolith, according to the retention time of the standard lysozyme.…”

mentioning

confidence: 99%

A Thermosensitive Monolithic Column as an Artificial Antibody for the On‐line Selective Separation of the Protein

Qin

Man

et al. 2010

Chemistry A European J

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The main objective of this study was to develop a new methodology for the preparation of a protein (antigen) that is a molecularly imprinted polymer (MIP, an artificial antibody) modified onto the surface of a silica skeleton in which the resulting stationary phase is thermosensitive. The silica monolithic skeleton with vinyl groups was synthesized in a stainless‐steel column by using a mild one‐step sol–gel process with two types of precursor: methyltrimethoxysilane (MTMS) and γ‐methacryloxypropyltrimethoxysilane (γ‐MAPS). Subsequently, three types of the thermosensitive protein MIP were anchored onto the surface of the silica skeleton to prepare the MIP monoliths, which were systematically investigated for back pressure and separation ability at different temperatures to establish good imprinting conditions. Under the optimized imprinting conditions, the chromatographic behavior of the thermosensitive MIP monolith exhibited strong retention ability for the lysozyme template (target antigen) in relation to the nonimprinting monolith (NIP monolith). The imprinting factor (IF) for lysozyme reached 3.48 at 20 °C. Moreover, this new type of artificial antibody displayed favorable binding characteristics for lysozyme over competitive proteins and was further evaluated to selectively separate lysozyme in a real sample by using an on‐line method. The run‐to‐run and column‐to‐column repeatability measurements of the thermosensitive MIP monoliths were also satisfactory.

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Affinity membrane chromatography: relationship of dye-ligand type to surface polarity and their effect on lysozyme separation and purification

Cited by 78 publications

References 38 publications

Dye Affinity Nanofiber Membrane for Adsorption of Lysozyme: Preparation and Performance Evaluation

Dye Affinity Nanofiber Membrane for Adsorption of Lysozyme: Preparation and Performance Evaluation

Purification of Egg White Lysozyme from Indonesian Kampung Chicken and Ducks

A Thermosensitive Monolithic Column as an Artificial Antibody for the On‐line Selective Separation of the Protein

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