INDUCTION OF IMMUNE RESPONSE AGAINST OESTRONE SULFATE PROTEIN CONJUGATFSIN RABBITS

Abstract

Development of immune response against estrone su lfate was evoked in rabbits. New estrone sulfate derivative was prepared and cross linked to bovine serum albumin with epitope ratio of 18 to be used as immunogen. Another estrone derivative was prepared and conjugated to rabbit gamma globulin with epitope ratio of 128 to elicit immune tolerance against cross reacting steroids. Immunization protocol comprised the use of dimetyl dioctadecyle ammonium bromide in comparison with heat killed Mycobacterium tuberculosis as adjuvants. Additionally, tetra 3H-estrone sulfate was prepared with 351tci/J..Lg specific activity for binding examination of antibodies. The itrununization protocol resulted in usable antibody titer (1:4900) and the immune tolerance technique gave high specific antisera with definite lower cross reactivity with estradiol and estriol sulfates (0.003 and <o.oool% respectively).Application of generated antibody in radioimmunoassay system was performed with bovine hormone free serum matrix. Standard curve constructed covering the range from 25-5000ng/50J..Ll was optimized for blocking agent 8-anilino-1-naphthalene sulfonic acid. Measurement of estrone su lfate in serum samples collected from a group of pregnant cows showed a grad u al increase of the steroid metabolite in blood with a peak value 24.82ng/ml blood at parturition.