Journal of agriculture & life science (농업생명과학연구)

Institute of Agriculture & Life Science, Gyeongsang National University (경상대학교 농업생명과학연구원)
권호 표지

Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocyte for Reducing Body Fat of Rat

흰쥐 체지방 감소를 위한 지방부위별 특이 다클론 항체 개발 연구

  • 최창원 (대구대학교 동물자원학과) ;
  • 박동우 (대구대학교 동물자원학과) ;
  • 조익환 (대구대학교 동물자원학과) ;
  • 오영균 (농촌진흥청 국립축산과학원) ;
  • 장선식 (농촌진흥청 국립축산과학원) ;
  • 최낙진 (전북대학교 동물소재공학과) ;
  • 감동근 (카길애그리퓨리나주식회사) ;
  • 백경훈 (영남대학교 생명공학부)
  • Received : 2012.02.07
  • Accepted : 2012.10.25
  • Published : 2012.10.31
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Abstract

Polyclonal antisera against regional (abdominal and subcutaneous) fats were developed to reduce body fat in rats. Isolation and culture of abdominal and subcutaneous adipocytes of rats were performed for analyzing lactate dehydrogenase (LDH) concentration. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas both of antisera against abdominal (AAb) and subcutaneous adipocyte plasma membrane proteins (SAb) had relatively strong reactivity till the level of 1:128,000 dilutions. Compared with regional fats, extremely low reactivities of AAb and SAb were detected with PMP of the organs (p<0.001). Both AAb and SAb were most strongly reacted with each adipocyte plasma memebrane proteins and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum based on LDH analysis. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte plasma membrane proteins are well developed and have safety in cross-reactivities with body organs.

본 연구는 가축 체지방 감소를 위한 수동면역학적 기법의 기초자료 확보를 위해 흰쥐 지방부위별(복강 및 피하지방) 특이 다클론 항체를 개발하고자 실시되었다. 흰쥐 부위별 지방 조직에서 지방세포를 분리하고 각각 primary 배양시킨 후 개발된 복강지방(AAb) 및 피하지방 특이 다클론 항체(SAb)를 주입한 뒤 media 내 lactate dehydrogenase(LDH) 방출 수준을 조사하였다. 희석배율 1:1,000배를 기준으로 비면역 혈청은 항원-항체 결합 반응이 거의 측정되지 않았고, AAb 및 SAb는 희석배율 1:128,000배까지 각각 항원-항체 반응이 감지되었다. 부위별 지방과 비교했을 때, AAb 및 SAb는 흰쥐 타 장기들과는 특이한 반응을 나타내지 않았다(p<0.001). 본 연구에서 개발한 두 항체들은 모두 항원으로 이용된 부위의 지방세포 PMP와 가장 높은 반응을 나타내었으며, 지방세포 배양을 통한 LDH 수준 검사에서는 AAb 및 SAb 모두 비면역혈청에 비해 유의적으로 높은 세포파괴가 일어났음을 확인할 수 있었다(p<0.01). 이상의 결과를 종합할 때 본 연구에서 개발된 AAb 및 SAb는 높은 역가, 타장기 안전성 및 in vitro 지방 감소 효과가 있었다고 생각된다.

Keywords

Acknowledgement

Supported by : 농촌진흥청

References

  1. Baek, K. H. 2003. Studies on the production of lean meats and the identification of Hanwoo (Korean Cattle) brand beef using immunological techniques. Ph.D. thesis. Yeungnam University, Gyeongsan, Korea.
  2. Baek, K. H., E. J. Kwon, T. H, Kwak, K. K. Jung, and C. B. Choi, 2000. In vitro cytotoxicity of polyclonal antibodies against proteins isolated from adipocyte plasma membrane of rats. J. Anim. Sci. & Technol. 42: 261-268.
  3. Baek, K. H., T. H. Kwak, Y. S. Oh, C. W. Choi, K. K. Jung, and C. B. Choi. 2005. Studies on the development and utilization of polyclonal antibodies against swine adipocyte plasma membrane proteins. J. Anim. Sci. & Technol. 47: 19-28. https://doi.org/10.5187/JAST.2005.47.1.019
  4. Choi, C. B. and S. R. Lee. 1996. Studies on the production of lean pork by immunological approach - Development of antibodies to porcine adipocyte plasma membrane. Korean J. Anim. Sci. 38: 369-374.
  5. Choi, C. B., M. J. Lee, and E. J. Kwon. 1997. Development of polyclonal antibody to adipocyte plasma membrane proteins isolated from Korean Naive Cattle. Korean J. Anim. Sci. 39: 669-674.
  6. Choi, C. B., M. J. Lee, and E. J. Kwon. 1998. Production of polyclonal antibodies specific to procine adipocyte plasma membrane proteins in sheep. Korean J. Biomed. Lab. Sci. 4: 57-63.
  7. Choi, C. W. 2011. Sera taken from aged Korean native steers increase adipocyte differentiation. J. Agr. Life Sci. 45: 85-92.
  8. Choi, C. W., K. H. Baek, C. B. Choi, Y. K. Oh, and S. K. Hong. 2010. Development of polyclonal antibodies to abdominal and subcutaneous adipocytes for producing fat-reduced high quality pork. Korean J. Food Sci. Ani. Resour. 30: 87-94. https://doi.org/10.5851/kosfa.2010.30.1.87
  9. Choi, C. W., Y. H. Kim, S. J. Kim, M. K. Song, E. K. Kwon, Y. K. Oh, S. G. Hong, S. H. Choi, and K. H. Baek. 2008. Development of polyclonal antibodies to abdominal and subcutaneous adipocytes for fat-reduced Hanwoo beef production. Korean J. Food Sci. Ani. Resour. 28: 651-659. https://doi.org/10.5851/kosfa.2008.28.5.651
  10. Cryer, A., B. R. Gray, and J. S. Woodhead. 1984. Study on the characterization of bovine adipocytes precursor cells and their differentiation, in vitro, using an indirect-labelled-second antibody cellular immunoassay. J. Develop. Physiol. 6: 159-176.
  11. Dong, J., D. P. Froman, and C. Y. Hu. 1990. Development and characterization of polyclonal antibodies against chicken adipocytes. Comp. Biochem. Physiol. 99: 195-198.
  12. Dulor, J. P., Y. Reyne, and J. Nougues. 1990. In vivo effects of a treatment with antibodies to adipocyte plasma membranes in the rabbit. Reprod. Nutr. Dev. 30: 49-58. https://doi.org/10.1051/rnd:19900104
  13. Duncan, D. B. 1955. Multiple range and multiple F test. Biometrics. 11: 1-42. https://doi.org/10.2307/3001478
  14. Flint, D. J., H. Coggrave, C. E. Futter, M. J. Gardner, and T. J. Clarke. 1986. Stimulatory and cytotoxic effects of an antiserum to adipocyte plasma membranes on adipose tissue metabolism in vitro and in vivo. Int. J. Obesity. 10: 69-77.
  15. Kestin, S., R. Kenndy, E. Tonner, M. Kiemam, A. Cryer, H. Griffin, S. Butterwith, S. Rhind, and D. J. Flint. 1993. Decreased fat content and increased lean in pigs treated with antibodies to adipocyte plasma membranes. J. Anim. Sci. 71: 1486-1494.
  16. Lafonton, M. and M. Berlan. 1984. Plasma membrane properties and receptors in while adipose tissue. In New Perspectives in Adipose Tissue: Structure, Function and Development. pp. 145-182. Cryer A. and R. L. R. Van. Eds. Butterworths, London. UK.
  17. Lo, H. F., T. R., August, U. A. Libeman, and I. S. Edelman. 1976. Dependence of renal $(Na^{+}^{+}K^{+})$-adenosine triphosphatase activity on thyroid status. J. Biol. Chem. 251: 7826-7833.
  18. Nassar, A. H. and C. Y. Hu. 1991. Antibodies to ovine adipocyte plasma membranes recognize tissue and species specific plasma membrane components. Comp. Biochem. Physiol. 98B: 361-367.
  19. Nassar, A. H. and C. Y. Hu. 1992. Characterization of polyclonal antibodies against ovine adipocyte plasma membranes. Int. J. Biochem. 24: 599-604. https://doi.org/10.1016/0020-711X(92)90333-V
  20. Panton, D., C. Futter, S. Kestin, and D. Flint. 1990. Increased growth and protein deposition in rats treated with antibodies to adipocytes. Am. J. Physiol. 258: E985-989.
  21. Paik, K. H., E. J. Kwon, T. H. Kwak, K. K. Jung, and C. B. Choi. 2000. In vitro cytotoxicity of polyclonal antibodies against proteins isloated from adipocyte plasma membrane. Korean J. Anim. Sci. & Technol. 42: 261-268.
  22. SAS. 2002. SAS User's Guide, Statistics, Version 9.1 Ed. SAS Inst., Inc., Cary, NC.
  23. Slavin, B. G. 1987. The ultrastructure of adipocytes. In Biology of the adipocyte. pp. 55-85. Hausman G. J. and R. Martin. Eds. Van Nostrand Reinhold, New York. USA.
  24. Vernon, R. G. 1986. The growth and metabolism of adipocytes. In Control and manipulation of animal growth. pp. 67-83. Buttery, P. J., D. B. Lindsay, and N. B. Hymes. Eds. Butterworths, London. UK.