Lavayre J, Leng M
Biochimie. 1977;59(1):33-42. doi: 10.1016/s0300-9084(77)80083-7.
Antibodies to adenosine were elicited in rabbits by immunization with bovine serum albumin-adenosine conjugate. The antibodies were purified and fractionated on two affinity columns (Sepharose-oligo(A) and Sepharose-AMP). Two families of antibodies have been obtained. The antibodies purified on the Sepharose-oligo(A) column react with poly(A) while those purified on the Sepharose-AMP column do not, as shown by gel diffusion. The association constants for the binding of Fab fragments or IgG purified on the Sepharose-oligo(A) column and several haptens were deduced from dialysis equilibrium, fluorescence quenching and displacement of AMP-fluorescein conjugate. The antibodies mainly recognize adenine, and the ribose or the phosphate group of (or AMP derivatives) do not play a critical role in the interaction. Thermodynamic parameters for adenosine-Fab fragments complexes have been determined deltaH degrees = 16 kcal/mole and deltaS degrees = - 15 cal. degree-1 mole-1. Circular dichroism studies indicate that about three nucleotide residues penetrate the binding site of Fab fragments.
通过用牛血清白蛋白 - 腺苷偶联物免疫兔子，引发了针对腺苷的抗体。抗体经纯化后在两个亲和柱（琼脂糖 - 寡聚（A）和琼脂糖 - AMP）上进行分离。已获得两类抗体。如凝胶扩散所示，在琼脂糖 - 寡聚（A）柱上纯化的抗体与聚（A）反应，而在琼脂糖 - AMP柱上纯化的抗体则不反应。从透析平衡、荧光猝灭以及AMP - 荧光素偶联物的置换反应中推导出在琼脂糖 - 寡聚（A）柱上纯化的Fab片段或IgG与几种半抗原结合的缔合常数。这些抗体主要识别腺嘌呤，并且（或AMP衍生物的）核糖或磷酸基团在相互作用中不起关键作用。已确定腺苷 - Fab片段复合物的热力学参数：ΔH° = 16千卡/摩尔，ΔS° = - 15卡·度⁻¹·摩尔⁻¹。圆二色性研究表明，约三个核苷酸残基穿透Fab片段的结合位点。
