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Helicobacter pylori로부터 유래된 항원의 anti-H, pylori 항체에 관한 연구
Study on Anti-Helicobacter pylori Antibody of Sparated Antigen from H. pylori

박창호   (대구바이오산업지원센터CC0126969  ); 배만종   (한의대학교 한방바이오식품과학과UU0000372  );
  • 초록

    축산물을 이용하여 생산된 특이항체는 세균성감염에 의한 설사병 치료와 충치예방에 효과가 있고, 특히 계란을 이용한 IgY (Immunoglobulin Yolk)는 비교적 산과 열에 안정하다는 실험결과가 보고되어[22] 있다. 식품분야에 특이항체를 식품소재로 산업화에 활용한 빈도는 아직 미미한 상태에 있으나, 최근 면역학, 단백질공학, 생명공학 등의 발전과 기술 향상으로 식품 소제로써의 활용성이 점차 높아질 것으로 기대된다. 본 연구에서는 H. pylori의 감염을 예방하고 치료보조제로 사용할 목적으로 포유동물을 통한 피동면역용 항체를 생산하고자 하였다. H. pylori로부터 항원을 분리하고, 분리된 항원에 대한 항체생산 및 H. pylori의 응집정도를 알아보았다. H. pylori로부터 분리된 주요 항원 단백질은 WC, OMP, crude urease, LPS 각각 12개, 7개, 3개, 1개 종류의 band를 확인할 수 있었다. 분리된 항원의 IgG 항체 생성은 동일한 항원농도인 $20{\mu}g/100{\mu}l$ 에서 각각 WC (L) $77.9{\pm}6.4{\mu}g/ml$ , OMP $84.9{\pm}6.4{\mu}g/ml$ , crude urease $123.8{\pm}2.9{\mu}g/ml$ 로 crude urease 항원이 가장 많은 것으로 나타났다. 그리고 IgA 항체 생성은 WC (L) $2.5{\pm}0.32{\mu}g/ml$ , OMP $2.0{\pm}0.43{\mu}g/ml$ , crude urease $1.3{\pm}0.25{\mu}g/ml$ 로 IgA 항체 생성은 WC (L) 항원이 가장 많은 것으로 나타났다 Western blotting을 통하여 항원 단백질의 면역원성 알아본 결과 WC 10종류, OMP 6종류, crude urease 3종류의 주요 항원성 물질을 확인할 수 있었다. 항체의 H. pylori에 대한 응집정도를 나타내는 응집가는 anti-WC (H), anti-WC (L), anti-OMP, anti-crude urease 항혈청에서 각각 $2^5,\;2^5,\;2^6\;and\;2^7$ 으로 나타났으며, 상대적으로 anti-crude urease 항혈청이 가장 높은 응집가를 나타내었다. 각 항원에 의해 생성된 항혈청의 urease활성 억제에 대한 흡광도(OD=550 nm)는 WC (H) $0.14{\pm}0.01$ , WC (L) $0.16{\pm}0.01$ , OMP $0.18{\pm}0.03$ , Urease $0.18{\pm}0.04$ 로 대조구 $0.26{\pm}0.02$ 와 비교할 때 유의적인 억제효과가 있는 것으로 나타났다. 이상의 결과를 종합해 볼 때 H. pylori 항원의 분리와 분리된 항원의 항체 생성능, H. pylori의 응집가, urease 활성억제측면에서 WC 및 crude urease항원 모두 높은 항체 역가를 나타내었다.


    This study has been carried out to secretion antibodies for the purpose of preventing the infection of Helicobacter pylori and using them as a supplement for treatment. This experiments have been separated antigens from H. pylori and observed into antibody production and the agglutination of H. pylori for the separated antigens. As major antigenic proteins separated from H. pylori, the following could be verified: 12 kinds of band for whole cell (WC), seven kinds of band for outer membrane protein (OMP), three kinds of band for crude urease, and one kind of band for lipopolysaccharide (LPS). The IgG anti-H. pylori antibody of separated antigens showed $77.9{\pm}6.4{\mu}g/ml$ for we (L), $84.9{\pm}6.4{\mu}g/ml$ for OMP, and $123.8{\pm}2.9{\mu}g/ml$ for crude urease, at the same antigen concentration of $20{\mu}g/100ull$ , which showed the most at the crude urease. And it turned out that the IgA antibodies were generated with $2.5{\pm}0.32{\mu}g/ml$ for WC (L), $2.0{\pm}0.43{\mu}g/ml$ for OMP, and $1.3{\pm}0.25{\mu}g/ml$ for crude urease, which demonstrated the most for WC (L) antigens. As a result of verifying the immunogenecity of antigenic protein through the Western blotting, major antigenic substances could be confirmed as follows: 10 kinds for WC, six kinds for OMP and three kinds for crude urease. The agglutination values on the H. pylori of the antibody were $2^5,\;2^5,\;2^6\;and\;2^7$ at the antigen serums of anti-WC (H), anti-WC (L), anti-OMP and anti-crude urease, respectively, which indicated the highest for the antigen serum of anti-crude urease. The urease activation-inhibiting absorbance of antigen serum created by each antigen was $0.14{\pm}0.01$ for WC (H), $0.16{\pm}0.01$ for WC (L), $0.18{\pm}0.03$ for OMP, and $0.18{\pm}0.04$ for urease, demonstrating a significant inhibiting effect, compared with $0.26{\pm}0.02$ of the control group.


  • 주제어

    Helicobacter pylori .   anti-H.   pylori antibody .   whole cell .   outer membrane protein .   lipopolysaccharide .   urease.  

  • 참고문헌 (29)

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