Rangel PLE, Perera MG, Lassala IA, Zarco QLA

Language: English/Spanish
References: 27
Page: 289-295
PDF size: 73.07 Kb.

ABSTRACT

Production of anti-cortisol antibodies was induced in laying hens. A conjugate of cortisol with bovine serum albumin (BSA) was mixed with Freund’s complete adjuvant and phisiological saline solution. One mg of the steroid was injected, followed by six more injections of 0.5 mg administered every two weeks. During egg collection, a serum titre average of 39.4% at a work dilution of 1:1000 was obtained. In order to develop a liquid phase radioimmunoassay (RIA), the anti-cortisol antibodies were recovered from the egg yolk, and then purified. Antibodies showed high specificity to cortisol, less than 0.1% cross-reaction with androstenedione, estradiol, 17-hidroxyprogesterone and testosterone with a sensitivity of 339 pg/ml. Accuracy of the RIA was appropriate (less than 15% interassay variation coefficient). The final yield was 594,552.63 mg of protein which can be used to process 95,000,000 tubes. It was concluded that the egg yolk of immunized hens is a good source of anti-cortisol antibodies, and that it can be used as an alternative to rabbit serum.

REFERENCES

1. Zarrow MX, Yochim JM, McCarthy JL. Experimental endocrinology. A sourcebook of basic techniques. New York: Academic Press, 1964.

2. Abraham G, Garza R. Radioimmunoassay of steroids. In: Abraham GE, editor. Handbook of radioimmunoassay. New York: Morton K. Schwartz, 1977.

3. Patterson R, Youngner JL, Weigle WO, Dixon FJ. Antibody production and transfer to egg yolk in chickens. J Immunol 1962;89:272-278.

4. Bauwens R, Devos M, Kint J, De Leenher A. Chicken egg yolk and rabbit serum compared as sources of antibody for radioimmunoassay of 1,25-dihydroxyvitamin D in serum plasma. Clin Chem 1988;34:2153-2154.

5. Gassmann M, Thömmes P, Weiser T, Hübscher U. Efficient production of chicken egg yolk antibodies against a conserved mammalian protein. FASEB 1990;4:2528-2532.

6. Otani H, Matsumoto K, Saeki A, Hosono A. Comparative studies on properties of hen egg yolk IgY and rabbit serum IgG antibodies. Lebensm Wiss Technol 1991;24:152-158.

7. Schade R, Pfister C, Halatsch R, Henklein P. Policlonal IgY antibodies from chicken egg yolk: an alternative to the production of mammalian IgG type antibodies in rabbits. ATLA 1991;19:403-419.

8. Svendsen L, Crowley A, Ostergaard H, Stodulski G, Hau J. Development and comparison of purification strategies for chicken antibodies from egg yolk. Lab Anim Sci 1995;45:89-93.

9. Svendsen L, Crowley A, Stodulski G, Hau J. Antibody production in rabbits and chickens immunized with human IgG. A comparison of titre and avidity development in rabbit serum, chicken serum and egg yolk using three different adyuvants. J Immunol Methods 1996;191:113-120.

10. Larsson A, Sjöquist J. Chicken antibodies: a tool to avoid false positive results by rheumatoid factor in the latex fixation tests. J Immunol Methods 1988;108:205-210.

11. Benson HN, Brumfield HP, Pomeroy BS. Requirement of avian C1 for fixation of Guinea pig complement by avian antibody-antigen complex. J Immunol 1961;87:63-68.

12. Montes P, Murcia M, Zarco Q. Producción de anticuerpos antiprogesterona a partir de yema de huevo de gallinas y del suero sanguíneo de conejos, para ser utilizados en radioinmunoanálisis. Vet Méx 1994;25:117-125.

13. Ermeling B, Steffen E, Fish R, Hook R. Evaluation of subcutaneous chambers as alternative to conventional methods of antibody production in chickens. Lab Anim Sci 1992;42:402-407.

14. IAEA. Laboratory training manual on radioimmunoassay in animal reproduction. Technical reports series No. 233. Vienna, Austria: FAO, International Atomic Energy Agency, 1984.

15. Patterson R, Youngner JL, Weigle WO, Dixon FJ. The metabolism of serum proteins in the hen and chick and secretion of serum proteins by the ovary of the hen. J Gen Physiol 1962b;45:501-513.

16. Garvey S, Cremer E, Sussdort H. Methods in immunology 3rd ed. London: WA Benjaminton, 1979.

17. Nicoll SC, Licht P. Evolutionary biology of prolactin and somatotropins. II. Electrophoretic comparison of tetrapod somatotropins. Gen Comp Endocrinol 1971;17:490-507.

18. Munro C, Stabenfeldt G. Development of a microtitre plate enzyme immunoassay for the determination of progesterone. J Endocrinol 1984;101:41-49.

19. Bedolla TN, Ulloa-Aguirre A, Landeros VJ, Pérez- Palacios G. Análisis de datos y control de calidad en el radioinmunoanálisis. I. Guía para la evaluación de resultados. Rev Invest Clin 1984;36:179-192.

20. Amyx HL. Control of animal pain and distress in antibody production and infectious disease studies. J Am Vet Assoc 1987;191:1287-1293.

21. Broderson JR. A retrospective review of lessons associated with the use of Freund’s adyuvant. Lab Anim Sci 1989;39:400-405.

22. Rangel PL. Comparación de dos fuentes biológicas para producir anticuerpos destinados al desarrollo de métodos de radioinmunoanálisis para determinar los niveles plasmáticos de testosterona y cortisol (tesis de maestría). México (DF) México: Facultad de Medicina Veterinaria y Zootecnia. UNAM, 1998.

23. Tizard I. Inmunología veterinaria. 3ª ed. México (DF): Interamericana, 1989.

24. Zar JH. Bioestatistical analysis. 3rd ed. Newark (NJ): Prentice Hall, New Jersey, 1996.

25. Russell WMS, Burch RL. The principles of humane experimental technique. South Mimms (UK): London Methuen & Co. Ltd., 1992.

26. Hsu WH, Crump MH. Glándula adrenal. En: McDonald L, editor. Endocrinología veterinaria y reproducción. 4ª ed. México (DF): Interamericana, 1989.

27. Libertun C. Radioinmunoanálisis. Fundamentos y aplicaciones. Buenos Aires, Argentina: López Libreros Editores,1980.