한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제24권4호
  • /
  • Pages.472-477
  • /
  • 1996
  • /
  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

임신부 뇨로부터 정제된 인간 상피세포 증식 인자 유사체의 in vitro bioassay 및 특성

  • 박세철 ((주)유한양행 중앙연구소) ;
  • 전재현 ((주)유한양행 중앙연구소) ;
  • 남정현 ((주)유한양행 중앙연구소) ;
  • 권태종 (건국대학교 미생물공학과) ;
  • 고인영 ((주)유한양행 중앙연구소) ;
  • 유광현 ((주)유한양행 중앙연구소)
  • 발행 : 1996.08.01
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초록

Natural human epidermal growth factor (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C$_{18}$ HPLC system. Following characterization by Western blot analysis and double immu- nodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporatioin of 5-bromo-2'-deoxy uridine (BrdU), a non-radioactive alternative for [$^{3}$H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1 - 10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others. Based on the results from the HPLC analysis spiked with recombinant human epidermal growth factor (rhEGF) and amino acid sequencing, we concluded that the second fraction was nhEGF and the other three fractions were the derivatives of nhEGF. In addition, the proportion of nhEGF was approximately 46% is compared with that of the other three derivatives.

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참고문헌

  1. J. Clin. Endo. Meta. v.66 Epidermal growth facor in urine of nonpregnant women and pregnant women throghout pregnancy and at delivery Hofman,G.;E.C.V.Rao;M.J.Brown;L.F.Murray;G.S.Schultz
  2. Hoppe-Seyler's Z. Physiol. Chem. v.356 The isolation of the urogastrones-inhibitors of gastric acid secretion from human urine Gregory,H.;I.R.Willshire
  3. Science v.189 Epidermal growth factor: Identification of a new hormone in urine Stanley,C.;N.O.David
  4. J. Invest. Derma. v.71 The regulation of cell proliferation: Advances in the biology and mechanism of action of epidermal growth factor Carpenter,G.
  5. Int. J. Petide protein Res. v.9 The primary structure of human urogastrone Gregory,H.;B.M.Preston
  6. Ann. Rew. Biochem. v.48 Epidermal growth factor Carpenter,G.
  7. Gastroenterology v.77 The idenification of urogastrone in serum, saliva and gastric juice Gregory,H.;S.Walsh;C.R.Hopkins
  8. Obstetries and Gynecology v.68 Peritoneal fluid, estrogen, progesterone, and epidermal growth factor concentrations in patients with and without endometriosis Leon,F.D.;R.Vijayakumar;M.Brown;C.H.Rao;M.A.Yussman;G.Schultza
  9. J. Clin. Endo. Metab. v.66 Epidermal growth factor in urine of nonpregnant women and pregnant women throughout pregnancy and at delivery Hofmann,G.E.;C.V.Rao;M.J.Brown;L.F.Murray;G.S.Schulta;T.A.Siddiqi
  10. Life Sciences v.41 Human epidermal growth factor concentrations in urine, but not in saliva and serum, depend on thyroid state Mattila,A.;L.J.perheentupa;J.Salmi;L.Viinikka
  11. Proc. Natl. Acad. Sci. USA. v.82 Sustained release of epidermal growth factor accelerated wound repair Buckly,A.;J.M.Davidson;C.D.Kamerth;T.B.Wolt;S.C.Woodward
  12. J. Lab. Clin. Med. v.108 Acceleration of wound healing by recombinant human urogastrone Franklin,T.J.;H.Gregory;W.P.Morris
  13. Chem. Pharm. Bull. v.33 Heterogeneity of human epidermal growth factor/urogastrone from human urine Nishimuro,S.;T.Nishimuro;N.Ulchida;H.Shirono;H.Hiratani
  14. J. Biol. Chem. v.247 Epidermal growth factor and a new derivative Savage,C.R.;C.Cohen
  15. Chromatographia v.27 I. HPLC analysis of human epidermal growth factor using immunoaffinity precolumn.Ⅱ. Determination of hEGFs in biological fluids Hayashi,T.;S.Sakamoto;I.H.Yoshida
  16. J. Biol. Chem. v.98 Preparation of extracts of the anterior pituitary-like substance of urine of pregnancy Philip,A.K.;A.D.Edward
  17. Nature v.227 Cleavage of structure proteins during the assembly of the head of bacteriophage Laemmli,U.k.
  18. Proc. Natl. Acad. Sci. v.76 Electrophoretic transfer of proteins from polyacrylamidegels to nitrocellulose sheets: procedure and some applications Towbin,H.T.;T.Stachelin;J.Gordon
  19. Anal. Biochem. v.111 Human epidernal growth factor/urogastrone: Rapid purification procedure and partial characterization Savage,C.R.;R.Happer
  20. Eur. J. Biochem. v.166 Purification of human growth factor (urogastrone) from urine Taspar,J.M.;P.Granchimont
  21. Archives of Biochem. Biophysics. v.240 Purification and characterization of epidermal growth factor and epidermal growth factor fragments from large volumes of human urine Charles,D.M.;J.L.Thomas;N.O.David
  22. J. Immunol. Methods. v.78 Single-step monoclonal affinity purification of human urogastrone from urine Hissey,P.H.;K.J.Thompson
  23. Anal. Saiences. v.4 Determination of epidermal growth factors in human urine by high performance liquid chromatography using anti-hEGF antibody precolumn Hayashi,T.;S.Sakamoto;T.Fuwa;I.Wada;H.Yoshida
  24. Differentiantion v.29 Biological effects of biosynthetic human EGF on the growth of mammalian cells in vitro Nakagawa,S.;A.Yoshida;Y.Hirao;S.Kasuga;T.Fuwa
  25. Science v.218 Monoclonal antibody to 5-Bromo-and 5-Iododeoxyuridine: a new reagent for detection of DNA replication Gratzner,H.G.
  26. Anal. Biochem. v.153 A biological assay for epidermal growth factor and related polypeptides Carpenter,G.;J.Zendequi
  27. Adv. Metab. Disord. v.8 Interaction of epidermal growth factor with cultured fibroblasts Stanley.C.;C.Graham;J.L.Kenneth
  28. J. Biol. Chem. v.270 Extacellular conversion of epidermal growth tactor (EGF) to des-Arg53-EGF by carboxypeptidase M McGwire.G.B.;R.A.Skidgel