A novel method for producing anti-peptide antibodies. Production of site-specific antibodies to the T cell antigen receptor beta-chain.
A novel method for producing anti-peptide antibodies. Production of site-specific antibodies to the T cell antigen receptor beta-chain.
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发表时间：
1988-02
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通讯作者：
D. Posnett;H. McGrath;J. Tam
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作者：
D. Posnett;H. McGrath;J. Tam
用于产生针对蛋白质的位点特异性抗体的肽抗原在疫苗的开发中是令人感兴趣的。需要将它们缀合至载体蛋白以获得最佳免疫原性导致许多问题，包括对载体的可能的免疫应答。在这里，我们描述了一种新的方法合成的免疫原性肽抗原，称为多抗原肽（MAP），这可能会使需要一个载体蛋白过时。选择源自人T细胞抗原受体β链恒定区的14个残基序列，并将该肽直接合成到分支赖氨酸核心上，其中8个拷贝的14个残基肽通过羧基末端氨基酸连接到核心上。该结构的分子量为13，422，其中仅7%代表核心的赖氨酸残基。八聚体MAP在小鼠和兔中具有高度免疫原性，允许产生多克隆和单克隆抗体。这些抗体中的大多数与其单体形式以及其八聚体形式的肽反应。此外，抗体与完整的β链蛋白反应。被抗体识别的肽的抗原决定簇包括连续决定簇和构象决定簇。八聚体MAP的NH 2-末端残基似乎是最具免疫原性的。没有针对中央赖氨酸核心的抗体。这种直接合成聚合肽的方法提供了免疫前肽的构象和数量的准确知识，而当肽与载体缀合时通常不是这种情况。该方法是通用的，因为存在合成具有16或32个肽臂的MAP或合成含有两种不同肽的聚合物的可能性。
Peptide antigens used to generate site-specific antibodies to proteins are of interest in the development of vaccines. The need to conjugate them to a carrier protein for optimal immunogenicity results in a number of problems including a possible immune response to the carrier. Here we describe a new method of synthesizing an immunogenic peptide antigen, referred to as multiple antigenic peptide (MAP), which may render the need for a carrier protein obsolete. A 14-residue sequence derived from the human T cell antigen receptor beta-chain constant region was selected, and the peptide was synthesized directly onto a branching lysine core with 8 copies of the 14-residue peptide linked to the core by the COOH-terminal amino acid. The molecular weight of this structure was 13,422 of which only 7% represents the lysine residues of the core. The octameric MAP was highly immunogenic in mice and rabbits, allowing production of polyclonal and monoclonal antibodies. The majority of these antibodies reacted with the peptide in its monomeric form as well as its octameric form. Moreover, the antibodies reacted with the intact beta-chain protein. The antigenic determinants of the peptide that were recognized by the antibodies included continuous determinants and conformational determinants. The NH2-terminal residues of the octameric MAP appeared to be most immunogenic. There were no antibodies to the central lysine core. This method of direct synthesis of a polymeric peptide provides accurate knowledge of the conformation and quantity of the peptide prior to immunization, which is usually not the case when peptides are conjugated to carriers. The method is versatile because the possibility exists to synthesize MAP with 16 or 32 peptide arms or to synthesize polymers containing two different peptides.