Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white

Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white

Fractionation of individual chicken egg white (CEW) proteins such as conalbumin (CNL), ovalbumin (OVL) and lysozyme (LYS) benefits the food, pharmaceuticals or nutraceuticals industry. Although packed bed chromatography is commonly used for protein separation, it still has a few limitations that nee...

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Main Author: Nurul Shamsinar, Shahirin
Format: Thesis
Language:English
Published: 2015
Subjects:
TP Chemical technology
Online Access:http://umpir.ump.edu.my/id/eprint/11246/19/Development%20of%20multiple%20interactions%20mixed%20matrix%20membrane%20chromatography%20for%20protein%20fractionation%20in%20chicken%20egg%20white.pdf
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spelling my-ump-ir.112462021-11-02T03:43:52Z Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white 2015-01 Nurul Shamsinar, Shahirin TP Chemical technology Fractionation of individual chicken egg white (CEW) proteins such as conalbumin (CNL), ovalbumin (OVL) and lysozyme (LYS) benefits the food, pharmaceuticals or nutraceuticals industry. Although packed bed chromatography is commonly used for protein separation, it still has a few limitations that need to be overcome. Membrane chromatography, which uses adsorptive membrane, is one alternative to overcome the limitation of packed bed column chromatography. Mixed matrix membrane (MMM) preparation concept provides a simple method for the preparation of membrane chromatography. In the current study, several potential low cost cation resins were screened to bind LYS at different pH. At pH 7, Lewatit CNP105 was selected for the preparation of cation exchanger MMM in the first part of study. The Lewatit CNP105 – cation exchanger MMM gave a Langmuir adsorption isotherm with the constant of 223 mg LYS/g membrane for qm and 0.045 for kd for single LYS solution. In batch binding of CEW solution at pH 7, both positively charged CEW protein (LYS, pI 10.7) and negatively charged CEW proteins (CNL, pI 6.1 and OVL, pI 4.5) were found to bind onto cation exchanger MMM. However, CNL and OVL were loosely bound and washed out during the washing step. Almost pure LYS can be recovered during the elution step of cation exchanger MMM chromatography.Multiple interactions MMM that combined Lewatit CNP105 cation resin and Lewatit MP500 anion resin, weretested to bind single protein and CEW solution in batch adsorption. In single protein experiment, the binding capacity of LYS, CNL and OVL are 52.3521 mg LYS/g membrane, 91.0453 mg CNL/g membrane and 10.8491 mg OVL/g membrane, respectively. The binding capacity of protein decreased in real CEW solution except for OVL. The reduction of the amount of protein binding might be due to the competitive binding with other impurities in CEW solution. For flow through experiment of multiple interactions MMM, three elution protocols were tested which were isocratic salt elution, linear gradient salt elution and combination of pH 3 and salt elution. Although no single protein fraction was recovered from these elution strategies tested, these fractions can be selected for further purification or used based on their targeted protein composition. 2015-01 Thesis http://umpir.ump.edu.my/id/eprint/11246/ http://umpir.ump.edu.my/id/eprint/11246/19/Development%20of%20multiple%20interactions%20mixed%20matrix%20membrane%20chromatography%20for%20protein%20fractionation%20in%20chicken%20egg%20white.pdf pdf en public masters Universiti Malaysia Pahang Faculty of Chemical & Natural Resources Engineering
institution Universiti Malaysia Pahang Al-Sultan Abdullah
collection UMPSA Institutional Repository
language English
topic TP Chemical technology
spellingShingle TP Chemical technology
Nurul Shamsinar, Shahirin
Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
description Fractionation of individual chicken egg white (CEW) proteins such as conalbumin (CNL), ovalbumin (OVL) and lysozyme (LYS) benefits the food, pharmaceuticals or nutraceuticals industry. Although packed bed chromatography is commonly used for protein separation, it still has a few limitations that need to be overcome. Membrane chromatography, which uses adsorptive membrane, is one alternative to overcome the limitation of packed bed column chromatography. Mixed matrix membrane (MMM) preparation concept provides a simple method for the preparation of membrane chromatography. In the current study, several potential low cost cation resins were screened to bind LYS at different pH. At pH 7, Lewatit CNP105 was selected for the preparation of cation exchanger MMM in the first part of study. The Lewatit CNP105 – cation exchanger MMM gave a Langmuir adsorption isotherm with the constant of 223 mg LYS/g membrane for qm and 0.045 for kd for single LYS solution. In batch binding of CEW solution at pH 7, both positively charged CEW protein (LYS, pI 10.7) and negatively charged CEW proteins (CNL, pI 6.1 and OVL, pI 4.5) were found to bind onto cation exchanger MMM. However, CNL and OVL were loosely bound and washed out during the washing step. Almost pure LYS can be recovered during the elution step of cation exchanger MMM chromatography.Multiple interactions MMM that combined Lewatit CNP105 cation resin and Lewatit MP500 anion resin, weretested to bind single protein and CEW solution in batch adsorption. In single protein experiment, the binding capacity of LYS, CNL and OVL are 52.3521 mg LYS/g membrane, 91.0453 mg CNL/g membrane and 10.8491 mg OVL/g membrane, respectively. The binding capacity of protein decreased in real CEW solution except for OVL. The reduction of the amount of protein binding might be due to the competitive binding with other impurities in CEW solution. For flow through experiment of multiple interactions MMM, three elution protocols were tested which were isocratic salt elution, linear gradient salt elution and combination of pH 3 and salt elution. Although no single protein fraction was recovered from these elution strategies tested, these fractions can be selected for further purification or used based on their targeted protein composition.
format Thesis
qualification_level Master's degree
author Nurul Shamsinar, Shahirin
author_facet Nurul Shamsinar, Shahirin
author_sort Nurul Shamsinar, Shahirin
title Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
title_short Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
title_full Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
title_fullStr Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
title_full_unstemmed Development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
title_sort development of multiple interactions mixed matrix membrane chromatography for protein fractionation in chicken egg white
granting_institution Universiti Malaysia Pahang
granting_department Faculty of Chemical & Natural Resources Engineering
publishDate 2015
url http://umpir.ump.edu.my/id/eprint/11246/19/Development%20of%20multiple%20interactions%20mixed%20matrix%20membrane%20chromatography%20for%20protein%20fractionation%20in%20chicken%20egg%20white.pdf
_version_ 1783731940164108288

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