In the last decades, protein engineering has developed particularly in biotechnology and pharmaceutical field. In particular, the engineered antibody subclass has arisen. The single chain diabody format (scDb), conjugating small size with antigen specificity, offers versatility representing a gold standard for a variety of applications, spacing from research to diagnostics and therapy. Along with such advantages, comes the challenge of optimizing their production, improving expression systems, purification procedures and stability. All such parameters are detrimental for protein production in general and above all for recombinant antibody expression, which has to be fine-tuned, choosing a proper protein-expression host and adjusting expression protocols accordingly. In the present paper, we present data regarding the production and purification of a single chain diabody directed against the macromolecular complex hERG1/β1 integrin. We focus on the expression of clones deriving from the transformation of Pichia pastoris yeast cells. In particular, we compare two different clones arose from two separate transformation processes, demonstrating that both are suitable for proper protein expression. Moreover, we have set up an expression protocol and compared the yields obtained using two purification machines: Akta Pure and Akta Start, with a positive outcome.

在过去的几十年中，蛋白质工程在生物技术和制药领域得到了特别的发展。特别是，工程抗体亚类已经出现。单链双抗体形式 (scDb) 将小尺寸与抗原特异性结合，提供多功能性，代表了各种应用的黄金标准，从研究到诊断和治疗。伴随这些优势而来的是优化其生产、改进表达系统、纯化程序和稳定性的挑战。所有这些参数通常对蛋白质生产有害，尤其对重组抗体表达有害，必须对其进行微调，选择合适的蛋白质表达宿主并相应地调整表达方案。在本文中，我们提供了关于生产和纯化针对大分子复合物 hERG1/β1 整联蛋白的单链双抗体的数据。我们专注于源自转化的克隆的表达毕赤酵母酵母细胞。特别是，我们比较了来自两个独立转化过程的两个不同克隆，证明两者都适合正确的蛋白质表达。此外，我们已经建立了一个表达协议，并比较了使用两台纯化机器获得的产量：Akta Pure 和 Akta Start，结果是积极的。