Purification of TGF-$\beta$1 from Bovine Colostrum

젖소 초유로부터 TGF-$\beta$1의 정제

  • 남명수 (충남대학교 동물자원학부) ;
  • 배형철 (충남대학교 동물자원학부) ;
  • 김평현 (강원대학교 미생물학과) ;
  • 김완섭 (강원대학교 축산가공학과) ;
  • 고준수 (강원대학교 축산가공학과)
  • Published : 2002.12.01

Abstract

Bovine colostrum contains various bio-functional Proteins. Especially, transforming: growth factor-${\beta}$1 (TGF-${\beta}$1) has a function in concerns with immune response. The purpose of this study was to establish the purification Processing of transforming growth factor-${\beta}$1(TGF-${\beta}$1). The highest concentration of TGF-${\beta}$1 was measured within 48 h after parturition in bovine colostrum using ELISA kit. Purification of TGF-${\beta}$1 from whey protein was carried out by the gel filtration, AF-heparin chromatography and AF-heparin rechromatography. After final purification step, TGF-${\beta}$1 with a molecular weight of 25 kD was obtained, and confirmed by silver staining and western blotting. Finally, TGF-${\beta}$1 was identified native form of 25 kD and reducing form of 12.5 kD by reducing agent.

TGF-$\beta$l은 여러 가지 생리활성 기능을 가지고 있기에 기능성식품 및 의약품 소재로 이용될 수 있다. 본 연구에서는 bovine colostrum milk로부터 TGF-$\beta$l을 분리 정제하기 위해 Cel-filtration chromatography, AF-heparin column chromatography 및 AF-heparin column rechromatography를 수행하여 TGF-$\beta$l을 정제하였다. 정제된 TGF-$\beta$1은 비환원조건하에서 전기영동을 수행하여 표준 TGF-$\beta$l과 같은 위치에 단일 band가 나타남으로 TGF-$\beta$l임을 확인하였다. 또한 환원 조건하에서 Western blot을 수행한 결과 TGF-$\beta$l 단일클론 항체와 결합하는 monomer 형태의 밴드를 확인하였다. 정제 TGF-$\beta$1의 회수율은 21%였다.

Keywords

References

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