Rezaei Javad, RajabiBazl Masoumeh, Ebrahimizadeh Walead, Dehbidi Gholamreza Rafiei, Hosseini Hossien
Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Protein Pept Lett. 2016;23(4):336-42. doi: 10.2174/0929866523666160115131403.
Prostate cancer (PCa) is the most frequently diagnosed cancer and the second most common cause of cancer related mortality in United States male population. ScFv fragments have different usefulness. For example they have small size, high perfusion rate, high yield of production and are non-immunogenic, thus they can be used for therapeutic purposes. In this project we used a synthetic human ScFv library for isolation of ScFv monoclonal antibodies against prostate specific membrane antigen. For this purpose, after five rounds of cell-panning, and also five rounds of antigen-panning with rPSMA specific anti- PSMA ScFv-phage particles were isolated. Phages with high affinity toward PSMA were selected and used for further analysis. Specificity and affinity of both ScFv to PSMA and LnCaP cell line examined by ELISA. Recombinant ScFv antibody isolated from cell-panning had higher specificity and affinity for both the antigen and LNCaP cell line. Our result demonstrated that ScFv antibody obtained by cell-panning can target PSMA antigen and cell lines.
前列腺癌（PCa）是美国男性人群中最常被诊断出的癌症，也是癌症相关死亡率的第二大常见原因。单链抗体片段（ScFv）具有不同的用途。例如，它们尺寸小、灌注率高、产量高且无免疫原性，因此可用于治疗目的。在本项目中，我们使用了一个合成的人ScFv文库来分离针对前列腺特异性膜抗原的ScFv单克隆抗体。为此，经过五轮细胞淘选，以及五轮抗原淘选后，分离出了具有rPSMA特异性的抗PSMA ScFv噬菌体颗粒。选择了对PSMA具有高亲和力的噬菌体并用于进一步分析。通过酶联免疫吸附测定（ELISA）检测了ScFv对PSMA和LnCaP细胞系的特异性和亲和力。从细胞淘选中分离出的重组ScFv抗体对抗原和LNCaP细胞系均具有更高的特异性和亲和力。我们的结果表明，通过细胞淘选获得的ScFv抗体可靶向PSMA抗原和细胞系。
