Relevance of oestrone presentation to the specificity of the elicited antisera, as revealed by affinity separation.
Polyclonal antisera raised against two different azobenzoyl-oestrone derivatives were analysed to investigate both the latency/intensity relationship of the immune response and the influence of antigen presentation on the specificity of the antisera elicited. Elongation of the azo-bridge of the hapten ([p(carboxyphenyl)-azo]-1,3,5[10]- oestratrien-3 ol-17 one) with a short aliphatic chain (4-amino-n-butyric acid) resulted in a marginal increase in the antibody yield, without affecting the time required to attain the maximum titre. The increased flexibility and mobility of the extended azo-bridge was shown to result in the appearance of antisera which cross-reacted with oestrogens with D ring structures different to that of oestrone. Antiserum fractionation by affinity chromatography through a stationary phase exposing the carrier protein determinants, as modified by the addition of the coupling bridge and the phenol ring, resulted in a reduction in its specificity. These findings are discussed with regard to the phenomena underlying the specificity of a polyclonal antiserum.
Podesta
A
A
University of Pisa, Department of Anatomy, Physiology and Animal Production, Italy.
Nieri
P
P
Luisi
M
M
Montagnoli
G
G
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D001391:Azo Compounds; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004970:Estrone / Q000032:analysis* / Q000276:immunology*; D006241:Haptens; D007106:Immune Sera / Q000302:isolation & purification*; D008297:Male; D011817:Rabbits / Q000276:immunology
D001391:Azo Compounds; D006241:Haptens; D007106:Immune Sera; D004970:Estrone