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Preliminary study on parameters of lysozyme purification using ion exchange chromatography

Wan Norlinda Roshana, Mohd Nawi (2006) Preliminary study on parameters of lysozyme purification using ion exchange chromatography. Faculty of Chemical & Natural Resources Engineering, Universiti Malaysia Pahang.


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Lysozyme is high demand bioproduct in market nowadays produced by protein purification process. Chromatography is a multicomponent separation technique which primarily used in the biotechnology sector especially in protein purification. The physical method of chromatography separation in which the components to be separated are distributed in stationary phase and mobile phase. One of the chromatography techniques that widely used is ion exchange chromatography (IEC) which based on the electrostatic interaction. The basic theory of the IEC separation process is depending on the isoelectric of the protein (pI). The pI of the protein is influence to determine the media of IEC whether anion or cation exchanger been used. In the experimental work, the automated liquid chromatography system from Amersham-Pharmacia Biosciences is designed for method development and research application whereas calibrated and controlled by a computer using UNICORN software. Preliminary study on the parameters is based on pH and salt concentration which to achieve the high purity of lysozyme. Some purification experiments were done to find the optimal operation condition to purify lysozyme from protein mixture by changing buffer pH and salt concentration in elution buffer to elute the interest protein. Based on the result and discussion; by manipulating pH value, the highest height detected at pH 8.5 which is 2.680. Using calibration curve, concentration of lysozyme is 0.0315 mg/ml. When NaCl concentration was manipulated, the concentration was optimizing at 0.3 M with is the result is 0.0284 mg/ml. As the conclusion, the optimum condition in purification step at pH buffer 8.5 and elution buffer at 0.3 M which is almost to the expected result refer to previous study.

Item Type: Undergraduates Project Papers
Uncontrolled Keywords: Proteins -- Purification
Subjects: Q Science > QP Physiology
Faculty/Division: Faculty of Chemical & Natural Resources Engineering
Depositing User: Rosfadilla Mohamad Zainun
Date Deposited: 15 Apr 2010 05:00
Last Modified: 03 Jun 2021 05:10
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