Leondiadis L, Livaniou E, Vassiliadou I, Ferderigos N, Ithakissios D S, Evangelatos G P
Institute of Radioisotopes and Radiodiagnostic Products, NCSR Demokritos, Athens, Greece.
Peptides. 1996;17(7):1091-6. doi: 10.1016/s0196-9781(96)00187-8.
We present theoretical and experimental data necessary for raising specific antibodies for thymosin beta 10, a 43-amino acid residues peptide occurring in human tissues. We postulate that thymosin beta 10 contains three major antigenic determinants (residues 2-8, 17-25, and 35-41). For antibody development, we synthesized the N-terminal fragment thymosin beta 10(1-16) as well as the C-terminal fragments thymosin beta 10(31-43) and thymosin beta 10(38-43), due to their putative antigenic properties and minimal structural similarity with the homologous peptide thymosin beta 4, which also occurs in humans. The putative antigenic determinant 17-25 is present in all beta-thymosins and was therefore not synthesized. All antisera raised against the above peptide fragments or the intact molecule of thymosin beta 10 were found capable of recognizing and binding synthetic or natural thymosin beta 10 with high specificity, showing minimal cross-reactivity with thymosin beta 4 isolated from bovine tissues or synthetic thymosin alpha 1. Due to its easy preparation and the highly specific affinity of the antibody raised against it for the intact peptide, the fragment thymosin beta 10(38-43) may be considered the antigen of choice for developing anti-thymosin beta 10 antibodies, which can eventually be applied to immunochemical studies.
我们提供了产生针对胸腺素β10（一种存在于人体组织中的43个氨基酸残基的肽）特异性抗体所需的理论和实验数据。我们推测胸腺素β10含有三个主要抗原决定簇（残基2 - 8、17 - 25和35 - 41）。为了开发抗体，我们合成了N端片段胸腺素β10(1 - 16)以及C端片段胸腺素β10(31 - 43)和胸腺素β10(38 - 43)，这是由于它们假定的抗原特性以及与同样存在于人类中的同源肽胸腺素β4的最小结构相似性。假定的抗原决定簇17 - 25存在于所有β - 胸腺素中，因此未合成。发现所有针对上述肽片段或胸腺素β10完整分子产生的抗血清都能够以高特异性识别并结合合成或天然的胸腺素β10，与从牛组织中分离的胸腺素β4或合成胸腺素α1的交叉反应最小。由于其易于制备以及针对它产生的抗体对完整肽具有高度特异性亲和力，胸腺素β10(38 - 43)片段可被视为开发抗胸腺素β10抗体的首选抗原，最终可应用于免疫化学研究。
