Experimental and Simulation Study of Lysozyme Separation in Cation Exchange Chromatography

양이온교환 크로마토그래피에서 Lysozyme 분리 실험과 전산모사

  • Kim, Jung-Ae (Department of Chemical Engineering, Chungnam National University) ;
  • Seong, Yeon-Kyeong (Department of Chemical Engineering, Chungnam National University) ;
  • Kim, In-Ho (Department of Chemical Engineering, Chungnam National University)
  • Published : 2006.06.28

Abstract

Lysozyme is an important antibacterial material, as effective food preservative. A number of lysozymes are found in nature such as egg white, where exists about 3.5% of egg proteins. In this study, carboxymethyl cation exchange chromatography has been used for separation of lysozyme. A simulation study by ASPEN was also performed for saving time and cost in chromatography purification experiments. Important parameters in experimental chromatography were sample loading amount, NaCl concentration, and pH of eluent. Simulation results were successfully fitted with chromatograms from experiments with change of parameters mentioned above.

이온교환 크로마토그래피에서 주요 변수로 샘플 투입량, 용출 NaCl 농도, 용출액 pH를 변화시켜 lysozyme 크로마토그라피 실험을 하였다. 그리고 Aspen Chromatography simulator를 이용하여 위의 변수 변화에 대한 모사된 크로마토그렘을 얻고 그 모사결과들을 실험 결과와 비교하였다. 용출 용액 NaCl의 농도가 높아질수록 단백질의 머무름 시간이 감소하며, 최대 피크의 높이가 높아졌다. pH가 증가함에 따라 최대 피크 높이가 증가하며, 분리 시간이 앞으로 이동하였다. 단백질의 주입량에 따라 분리 시간에 상관없이 최대 피크 높이만 비례적으로 증가하는 것을 확인하였다. 그리고 실험을 통해 얻어진 결과와 시뮬레이션 결과의 비교를 통해 같은 경향을 보임을 확인하였다.

Keywords

References

  1. Huh, Y. S., H. W. Kim, and I. H. Kim (2003), Purification of Lysozyme from Egg White by Multicycle Ion Exchange Chromatography, Korean J. Biotechnol. Bioeng. 18(2), 1-7
  2. Yao, S. J., Y. X. Guan, and L. H. Yu (2003), Adsorption Performance of Proteins to CM Sepharose FF and DEAE Sepharose FF Adsorbents , Korean J. Chem. Eng. 20(1), 93-98 https://doi.org/10.1007/BF02697191
  3. Lee, W. K and J. S. Ko (2003), Kinetic Model for the Simulation of Hen Egg White Lysozyme Adsorption at Solid/Water Interface, Korean J. Chem. Eng. 20(3), 549-553 https://doi.org/10.1007/BF02705564
  4. Jin, L. M., S. K. Han, D. K. Choi, and K. H. Row (2005), Nonlinear Adsorption Isotherm of Single and Multi-Components of 2'-Deoxyribonucleosides, Korea Chem. Eng. Res. 43(2), 230-235
  5. Aspen Chromatography User Manual (2002)
  6. Kim, H. W. and I. H. Kim (2003), Comparison of Lysozyme Purification from Egg White Between Ion Exchange Chromatography and Precipitation, HWAHAK KONGHAK 41(3), 332-336
  7. Kim, I. H. and J. T. Kim (1998), Pulse Response Test in Medium Pressure Protein Chromatography, Korea J. Chem. Eng. 15(6), 575-579 https://doi.org/10.1007/BF02698980