Verdoliva A, Ruvo M, Villain M, Cassani G, Fassina G
Protein Engineering, TECNOGEN S.C.p.A. Parco Scientifico, Piana di Monte Verna (CE), Italy.
Biochim Biophys Acta. 1995 Nov 15;1253(1):57-62. doi: 10.1016/0167-4838(95)00149-o.
Antibodies raised in rabbits against multimeric all-L peptides (MAP's) were first made monospecific by affinity chromatography on immobilized antigen columns and then tested for their ability to cross-react with topologically related variants of the parent antigen, where the chirality of each amino-acid residue (inverso derivatives), or the peptide sequence orientation (retro derivatives), was inverted, or where both modifications were simultaneously introduced (retro-inverso derivatives). Retro, inverso, and retro-inverso forms of the parent peptide were prepared, both in the linear as well as in the BSA-conjugated form, and found to cross-react to a significant extent with affinity purified polyclonal antibodies raised against the parent peptide. Peptide variants displayed similar dose-dependent inhibitory effects on the interaction between immobilized parent antigen and affinity purified antibodies. Analysis of molecular models of the peptide variants in the trans-configuration suggested that the topological equivalence of alternating side chains in the series of related peptides may be responsible for the observed cross-recognition, leading to the formation of similar recognition surfaces which could mimic the parent peptide antigenic structure.
用兔制备的针对多聚全L肽（MAP）的抗体，首先通过固定化抗原柱上的亲和层析使其具有单特异性，然后测试其与亲本抗原拓扑相关变体交叉反应的能力，这些变体中每个氨基酸残基的手性（反向衍生物）或肽序列方向（反向衍生物）被颠倒，或者同时引入两种修饰（反向-反向衍生物）。制备了亲本肽的反向、反向和反向-反向形式，既有线性形式也有与牛血清白蛋白（BSA）偶联的形式，发现它们与针对亲本肽产生的亲和纯化多克隆抗体有显著程度的交叉反应。肽变体对固定化亲本抗原与亲和纯化抗体之间的相互作用表现出类似的剂量依赖性抑制作用。对反式构型肽变体的分子模型分析表明，一系列相关肽中交替侧链的拓扑等效性可能是观察到的交叉识别的原因，导致形成类似的识别表面，该表面可模拟亲本肽的抗原结构。
