Tsuruta Lilian R, Hayashi Mirian A F, Konno Katsuhiro, Tambourgi Denise V, Assakura Marina T, Camargo Antonio C M, Sant'Anna Osvaldo A
Center for Applied Toxinology, Instituto Butantan, São Paulo, Brazil.
Anal Biochem. 2006 Jun 15;353(2):174-80. doi: 10.1016/j.ab.2006.03.018. Epub 2006 Mar 30.
Production of specific antibodies to haptens, especially antipeptides, without interference by carrier protein, is desirable. The bradykinin-potentiating peptides (BPPs) are a family of pyroglutamyl proline-rich oligopeptides with strong antihypertensive properties. In this work, the production of antibodies to BPPs by use of an efficient immunization protocol in mice genetically modified for the high antibody responsiveness (H(III) line) is described. Although it was possible to induce antibody production by single-dose administration of free BPPs, higher antibody titers were obtained in mice preimmunized with carrier protein before administration of peptides conjugated to this carrier. Interestingly, both mouse groups had a higher titer of IgG(1) than IgG(2a) isotypes, regardless of prior immunization with the carrier protein. However, a lower titer of IgG(2a) was observed in unprimed mice. A single band of about 27kDa corresponding to the BPP precursor protein was recognized by these antibodies in the cytosol of the Bothrops jararaca venom gland. This work proposes an efficient immunization protocol based on classic studies described for the hapten-carrier effect for generating specific antibodies against biologically active peptides.
在不受到载体蛋白干扰的情况下生产针对半抗原，尤其是抗肽的特异性抗体是很有必要的。缓激肽增强肽（BPPs）是一类富含焦谷氨酸 - 脯氨酸的寡肽家族，具有很强的降压特性。在这项工作中，描述了通过在具有高抗体反应性的基因改造小鼠（H(III)系）中使用高效免疫方案来生产针对BPPs的抗体。尽管通过单剂量给予游离BPPs可以诱导抗体产生，但在用与该载体偶联的肽给药前先用载体蛋白进行预免疫的小鼠中获得了更高的抗体滴度。有趣的是，无论之前是否用载体蛋白免疫，两个小鼠组的IgG(1)同种型滴度都高于IgG(2a)。然而，在未预免疫的小鼠中观察到较低的IgG(2a)滴度。这些抗体在Bothrops jararaca蛇毒腺的胞质溶胶中识别出一条约27kDa的对应于BPP前体蛋白的单一条带。这项工作基于对半抗原 - 载体效应描述的经典研究提出了一种高效免疫方案，用于产生针对生物活性肽的特异性抗体。
