Podesta A, Luisi M, Montagnoli G
National Council for Research, Institute of Biophysics, Roma, Italia.
J Steroid Biochem Mol Biol. 1991 Jul;39(1):63-71. doi: 10.1016/0960-0760(91)90014-v.
An immunosorbent technique was developed to attenuate cross-reactivity of a polyclonal antiserum against a 4(2) (rho-carboxyphenylazo)-1,3,5[10]-estratrien-3,16 alpha,17 beta-triol-bovine serum albumin conjugate. The chromatographic separation of antiserum through stationary phases having either rho(carboxymethyl)phenylazo-phenol or rho(carboxymethyl)-phenylazo-2-naphthol side residues reduced the antiserum avidity, while increasing the apparent antiserum affinity and decreasing the residual cross-reactivities against heterologous ligands. The highly specific antiserum obtained allowed the development of a competitive binding assay over an extended analytical range, which opens up the possibility of direct measurement of estriol from the early pregnancy to delivery. The significance of the attenuation of antiserum cross-reactions after affinity chromatography is discussed with reference to epitope-paratope interaction in the case of small endogenous molecules like estrogens.
开发了一种免疫吸附技术，以减弱多克隆抗血清对4(2)(对羧基苯偶氮)-1,3,5[10]-雌甾三烯-3,16α,17β-三醇-牛血清白蛋白偶联物的交叉反应性。通过具有对(羧甲基)苯偶氮苯酚或对(羧甲基)-苯偶氮-2-萘酚侧基的固定相对抗血清进行色谱分离，降低了抗血清的亲和力，同时增加了表观抗血清亲和力，并降低了对异源配体的残留交叉反应性。获得的高特异性抗血清使得能够在扩展的分析范围内开展竞争性结合测定，这为从早孕到分娩直接测量雌三醇开辟了可能性。结合雌激素等内源性小分子的表位-互补位相互作用，讨论了亲和色谱后抗血清交叉反应减弱的意义。
