KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Optimization of Chromatographic Separation of Lysozyme from Homogenate of Hen Egg White by Comparison of Breakthrough Behavior

파과분석(Breakthrough behavior) 비교에 의한 난백으로부터 라이소자임 크로마토그래피 분리 최적화

  • 김원경 (생명공학연구소 생물분리공정 Research Unit) ;
  • 정봉현 (생명공학연구소 생물분리공정 Research Unit)
  • Published : 1999.06.01
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Abstract

We have compared the breakthrough behavior of lysozyme contained in fresh han egg white on various cation exchagers, and the adsorbent, known by the trade name Cellufine C-200 (Amicon), has shown the best performance. The effects of ion strength, pH, and linear flow rate on the breakthrough behavior were examined using the Cellufine C-200 adsorbent. The optimal conductivity, pH and linear flow rate were determined from the breakthrough behavior and found to be 2.75 mS/cm, 7.0, and 0.635 cm/min, respectively.

본 연구에서는 난백에 포함된 라이소자임의 컬럼 크로마토그래피에서의 breakthrough behavior 실험을 통해 최적 크로마토그래피 분리조건을 결정하였다. 다양한 양이온 교환수지의 라이소자임 breakthrough behavior를 비교한 결과 Cellufine C-200(Amicon, USA)이 라이소자임 흡착에 가장 우수한 성능을 보였다. Cellufine C-200 수지에서 이온강도, pH, 선속도 등이 난백 라이소자임의 breakthrough behavior에 미치는 영향을 비교한 결과 conductivity 2.75 mS/cm, pH 7, 선속도 0.635cm/min의 최적 조건을 얻었다. 본 최적조건에서 prep 규모 컬럼 크로마토그래퍼를 이용 난백으로부터 라이소자임을 분리하였다. 정제된 라이소자임과 Sigma 라이소자임의 specific activity는 비슷하였으며 SDS-PAGE 분석 결과 본 연구에서 정제된 라이소자임이 Sigma 라이소자임보다 impurity 단백질들을 적게 포함하고 있음을 확인하였다.

Keywords

References

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