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首页> 外文期刊>International immunopharmacology >Generation and characterization of a specific single-chain antibody against DSPP as a prostate cancer biomarker: Involvement of bioinformatics-based design of novel epitopes
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Generation and characterization of a specific single-chain antibody against DSPP as a prostate cancer biomarker: Involvement of bioinformatics-based design of novel epitopes

机译：对DSPP作为前列腺癌的特定单链抗体的产生与表征生物标志物：新型表位的生物信息学的设计累及

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Isolation of specific single chain antibodies (scFvs) against key epitopes of cancer markers are applied for cancer immunotherapy and diagnosis. In this study following the prediction of the 3D structure of the DSP part of Dentin sialophosphoprotein (DSPP), the epitope was chosen using in silico programs. Panning process was applied to isolate specific human scFv against the epitope. PCR and DNA fingerprinting differentiated the specific clones, which were evaluated by phage ELISA. Following DNA sequencing, the 3D structure of isolated scFv was modeled and Docked on DSP. Results demonstrated the selection of a specific anti-DSPP scFv with 40% frequency, which reacted significantly with the predicted epitope and PCa patients' urines in ELISA tests (P-value < 0.05). The VH and VL of the isolated scFv were from VH1 and VL3 gene families with several amino acid changes in CDRs and FRs domains. The scFv tightly bound to the DSP epitope with the lowest energy level by hydrogen bonds, cation-pi, hydrophobic and ionic interactions demonstrating the specificity of Ag-Ab interactions. The anti-DSPP scFv selected in this study with significant specificity to DSPP antigen offers a promising new agent for both PCa early detection and treatment of cancers with DSPP expression.
机译：针对癌症免疫疗法和诊断的癌症标志物关键表位的特定单链抗体（SCFV）的分离。在该研究之后，在牙本质苷肽（DSPP）的DSP部分的3D结构的预测中，在硅程序中选择表位。将平移过程应用于将特异性人体SCFV分离对表位。 PCR和DNA指纹纹理分化了通过噬菌体ELISA评估的特定克隆。在DNA测序后，将隔离SCFV的3D结构进行建模并对接DSP。结果表明，选择具有40％频率的特定抗DSPP SCFV，其在ELISA测试中预测表位和PCA患者的尿液作出显着反应（p值<0.05）。分离的SCFV的VH和VL来自VH1和VL3基因家族，CDR和FRS结构域的几个氨基酸变化。通过氢键，阳离子-PI，疏水性和离子相互作用的能量水平最低的DSP表位紧密结合DSP表位，证明了AG-AB相互作用的特异性。本研究中选择的抗DSPP SCFV具有显着的DSPP抗原的特异性为PCA早期检测和治疗具有DSPP表达的癌症提供了有希望的新试剂。

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