A method of improving specific immune responses to smal immunogens, haptens, has been developed by changing the linkage between the hapten and carrier being used for immunization. High affinity antibodies to the hapten cotinine have been produced using this method. Antibodies to a glycated protein have also been developed, utilizing an immunogen which is composed of a glycated peptide mimic of the glycated peptide sequence which is the target epitope, wherein the peptide mimic is constructed to conformationally mimic the conformation of the peptide in the native protein, the peptide mimic contains no charged groups or other immunodominant group, and the peptide mimic is connected to a spacer sequence equivalent to a peptide spacer of between one and thirty amino acids in length, which serves to position the peptide epitope in a conformation that approximates its conformation in the native protein. In a further embodiment the peptide mimic and spacer are linked to a carrier molecule. This method has been used to produce an antibody to the glycated protein HbA1c, wherein the peptide mimic includes a valine modified by addition of a glucose molecule, an analog of Histidine which does not bear a charge in the immunizing structure, allows orientation of the peptide so that the immune response can be directed to the side of the peptide chain oriented oppositely to the ring, and is of a size that the conformation of the peptide mimics the conformation of the peptide in the native molecule, a leucine or an analog thereof which allows binding to an antibody preferentially recognizing Hb A1c such as 82D259, and a threonine or an analog thereof which allows binding to antibody number 82D259. In the example described below the histidine analog is 2-amino-3-flurylpropionyl, and the peptide is Fructosyl Val-2-amino-3-furanylproprionic acid-Leu-Pro-Pro-Glu-Glu-Tyr-Tyr-Cys.
通过改变杂合体与用于免疫的载体之间的联系,开发出了一种提高对小免疫原(杂合体)的特异性免疫反应的方法。利用这种方法产生了针对合蛋白
可替宁的高亲和力
抗体。还开发了糖化蛋白质
抗体,利用的免疫原由目标表位的糖化肽序列的糖化
肽模拟物组成,其中
肽模拟物的构造是为了在构象上模拟原生蛋白质中肽的构象、
多肽模拟物不含带电基团或其他免疫显性基团,
多肽模拟物与相当于一至三十个
氨基酸长度的
多肽间隔序列相连,该间隔序列的作用是将
多肽表位定位在近似于其在原生蛋白中的构象的位置。在另一个实施方案中,
多肽模拟物和间隔物与载体分子相连。这种方法已被用于生产糖化蛋白 HbA1c 的
抗体,其中的
肽模拟物包括一个通过添加
葡萄糖分子修饰的缬
氨酸,它是组
氨酸的类似物,在免疫结构中不带电,可使肽定向,从而使免疫反应指向肽链与环相反的一侧、组
氨酸或其类似物,可与优先识别 Hb A1c 的
抗体如 82D259 结合;苏
氨酸或其类似物,可与
抗体 82D259 结合。在下面描述的例子中,组
氨酸类似物是 2-
氨基-3-
呋喃丙酰基,肽是
果糖基 Val-2-
氨基-
3-呋喃丙酸-Leu-Pro-Pro-Glu-Glu-Tyr-Cys。