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首页> 外文期刊>Renewable Agriculture and Food Systems >Immunomagnetic separation of human myeloperoxidase using an antibody-mimicking peptide identified by phage display
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Immunomagnetic separation of human myeloperoxidase using an antibody-mimicking peptide identified by phage display

机译：用噬菌体展示鉴定的抗体模拟肽的免疫磁性分离

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Phage display biopanning is a powerful in vitro selection process for screening and identifying peptides that bind to a target protein of interest. With the aim of replacing antibodies in immuno-diagnostic applications, we identified peptides whose binding characteristics mimicked those of anti-human myeloperoxidase (hMPO), a biomarker for acute cardiac diseases. Based on ELISA results from four phage clones, we selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0.023, approximately 43-fold lower than that of the anti-hMPO antibody. The dissociation constant (K-d) was 57 nM, which was comparable to that of the native antibody (83 nM). Next, we biotinylated the peptide at its N-terminus and attached the biotinylated peptide to the surface of streptavidin-coated magnetic particles to assess its ability to selectively capture hMPO. The binding equilibrium data were similar to the previous analyses; specifically, around 0.021 mol peptide bound to 1 mol of hMPO. Antigen capture was found to be selective and to be relatively little influenced by the presence of human serum albumin (HSA), an abundant constituent of serum. Our work demonstrates the potential of immunomagnetic isolation to achieve selective capture of a low-concentration antigen from complex solutions such as serum. (C) 2016 Elsevier B.V. All rights reserved.
机译：噬菌体展示生物丙现是一种强大的体外选择方法，用于筛选和识别与目标靶蛋白结合的肽。随着替代免疫诊断应用中的抗体，我们鉴定了肽，其结合特征模仿抗人髓氧化酶（HMPO），一种用于急性心脏病的生物标志物。基于四个噬菌体克隆的ELISA，我们选择和化学合成了12-MEL肽（SyiepperHRHR）。石英晶微观和表面等离子体共振分析显示，合成肽的摩尔结合平衡比为0.023，比抗HMPO抗体低约43倍。解离常数（K-D）为57nm，其与天然抗体（83nm）的相当。接下来，我们将肽在其N-末端生物蛋白化并将生物素化的肽连接到链霉蛋白涂覆的磁性颗粒的表面中以评估其选择性捕获HMPO的能力。绑定平衡数据类似于先前的分析;具体而言，约0.021摩尔肽结合至1mol的HMPO。发现抗原捕获是选择性的，并且受人血清白蛋白（HSA）的存在，血清的丰富成分相对较少。我们的工作证明了免疫磁性隔离的潜力，以实现从血清如血清的复合溶液中选择性捕获低浓度抗原。 （c）2016年Elsevier B.v.保留所有权利。

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