KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Isolation of Peptides from Human Blood by RP-HPLC

RP-HPLC를 이용한 혈액에 포함된 펩타이드의 분리

  • 노경호 (초정밀생물분리기술센터, 인하대학교, 화학공학과) ;
  • 이승기 (초정밀생물분리기술센터, 인하대학교, 화학공학과) ;
  • Published : 2002.06.01
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Abstract

The biologically active peptides in human blood can adjust the functions of many physiological systems. The peptides in human blood were separated on the five steps of linear gradient-elution mode by RP-HPLC with UV detection. The size of commercially available $C_{18}$ chromatographic column was 4.60$\times$150 mm with particle size of 5 $\mu\textrm{m}$ and pore size of 100 $\AA$. The mobile phases used were water in 0.75% trifluoroacetic acids (TFA) and organic modifier of acetonitrile. The isolation methods suggested in this work for peptides from the blood were composed of the formation of immiscible liquid layers and precipitation by centrifuge and chemicals of sodium citrate and trichloroacetic acid (TCA). The some peptides were identified based on the retention times previously constructed database.

생리활성을 가지는 펩타이드에 관한 연구는 각종 질환의 진단, 예방 및 치료약으로서의 응용 가능성이 매우 높아서 현재 많은 연구가 진행 중이다. 펩타이드를 RP-HPLC에 의해 정량, 정성분석 함으로 다양한 연구 분야로의 확대를 모색해 볼 수 있다. 혈액 속에 있는 적혈구내의 펩타이드를 분리하기 위해 0.75% TFA가 포함된 물과 아세토나이트릴을 이동상과 선형적 구배용매 조성을 이용한 실험결과, 펩타이드의 peak 분리도는 상당히 우수하였다. 혈액을 채취한 날짜의 경과에 따라서 새로운 생리활성 물질이 만들어짐을 확인할 수 있었다. 이와 같은 현상은 적혈구내의 세포들이 산소를 계속적으로 소비함으로써 산소 부족현상 때문에 발생하는 것으로 알려져 있고 구조적 변화를 나타내는 생리활성 펩타이드를 정성적으로 확인함으로써 좀더 의학적인 분야로의 연구를 수 행할 수 있게 될 것이다.

Keywords

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