Sivaccumar Jwala P, Iaccarino Emanuela, Oliver Angela, Cantile Maria, Olimpieri Pierpaolo, Leonardi Antonio, Ruvo Menotti, Sandomenico Annamaria
Institute of Biostructures and Bioimaging, CNR, Via P. Castellino, 111, 80131 Naples, Italy.
Università degli Studi della Campania Luigi Vanvitelli, Via Vivaldi 43, 81100 Caserta, Italy.
Pharmaceuticals (Basel). 2023 Aug 10;16(8):1130. doi: 10.3390/ph16081130.
Drug development in recent years is increasingly focused on developing personalized treatments based on blocking molecules selective for therapeutic targets specifically present in individual patients. In this perspective, the specificity of therapeutic targets and blocking agents plays a crucial role. Monoclonal antibodies (mAbs) and their surrogates are increasingly used in this context thanks to their ability to bind therapeutic targets and to inhibit their activity or to transport bioactive molecules into the compartments in which the targets are expressed. Small antibody-like molecules, such as Fabs, are often used in certain clinical settings where small size and better tissue penetration are required. In the wake of this research trend, we developed a murine mAb (3D1) neutralizing the activity of Nodal, an oncofetal protein that is attracting an ever-increasing interest as a selective therapeutic target for several cancer types. Here, we report the preparation of a recombinant Fab of 3D1 that has been humanized through a computational approach starting from the sequence of the murine antibody. The Fab has been expressed in bacterial cells (1 mg/L bacterial culture), biochemically characterized in terms of stability and binding properties by circular dichroism and bio-layer interferometry techniques and tested in vitro on Nodal-positive cancer cells.
近年来，药物研发越来越侧重于开发个性化治疗方案，这些方案基于阻断对个体患者中特异性存在的治疗靶点具有选择性的分子。从这个角度来看，治疗靶点和阻断剂的特异性起着至关重要的作用。单克隆抗体（mAb）及其替代物由于能够结合治疗靶点并抑制其活性，或将生物活性分子转运到表达靶点的区室中，因此在这种情况下越来越多地被使用。小抗体样分子，如Fab片段，常用于某些需要小尺寸和更好组织穿透性的临床环境中。紧跟这一研究趋势，我们开发了一种鼠源单克隆抗体（3D1），它能够中和Nodal的活性，Nodal是一种癌胚蛋白，作为几种癌症类型的选择性治疗靶点，正吸引着越来越多的关注。在此，我们报告了一种3D1重组Fab片段的制备，该片段通过基于鼠源抗体序列的计算方法进行了人源化。该Fab片段已在细菌细胞中表达（每升细菌培养物1毫克），通过圆二色性和生物层干涉术技术对其稳定性和结合特性进行了生化表征，并在体外对Nodal阳性癌细胞进行了测试。
