噬菌体展示技术及其在肿瘤研究中的应用

噬菌体表面展示技术是一项特异性多肽或蛋白的筛选技术,它将随机序列的多肽或蛋白片段与噬菌体衣壳蛋白融合表达而呈现于病毒表面,被展示的多肽能保持相对独立的空间结构,使其能够与配体作用而达到模仿性筛选特异性分子表位,从而提供了高通量高效率的筛选系统。近年来噬菌体展示技术已广泛应用于肿瘤抗原抗体库的建立、单克隆抗体制备、多肽筛选、疫苗研制、肿瘤相关抗原筛选和抗原表位研究、药物设计、癌症检测和诊断、基因治疗及细胞信号转导研究等。就近年来噬菌体展示技术在肿瘤相关研究中的运用作以综述。     

噬菌体展示库筛选细胞特异分子的进展

介绍噬菌体展示技术的原理和发展,尤其是噬菌体展示技术在筛选细胞特异分子的策略方面的进展。该技术通过20年的发展已成为一种研究抗原一抗体作用、蛋白质相互作用、蛋白一药物相互作用甚至蛋白质一核酸作用的分析手段,但涉及到以完整细胞、器官或组织等复杂的生物活性分子表面为靶标则筛选效果尚不理想。关键是要减少噬菌体展示分子与靶标的非特异性结合,利用更为严格的经过改进的筛选策略。该技术的优势预示着它将广泛被应用于基础理论和研究实践中。     

噬菌体表面显示技术在生物医学研究中的应用

利用噬菌体表面显示技术,将蛋白质或短肽分子在噬菌体表面表达,可以对抗原表位及蛋白质的相互作用位点进行定位与模拟,并用于蛋白质的体外亲和力优化以及新基因的克隆,在小分子药物的设计和筛选中也具有重要的作用。环肽结构的设计以及筛选策略和方法的改进,逐步推动着噬菌体表面显示技术在免疫学、基因工程药物及基础生物医学研究等研究领域中的成功应用     

噬菌体表面展示肽库技术在病毒研究中的应用

噬菌体表面展示肽库技术是近年来发展起来的用丝状噬菌体展示外源肽的一项新技术,被广泛应用于分子识别的各个领域。本文就该技术在病毒受体、病毒抗原表位、病毒抗体以及病毒疫苗和病毒性疾病的诊断等央电荷和一简要概述。     

从七肽噬菌体展示库中筛选内毒素结合蛋白质配基

目的:研究从噬茵体展示库中筛选内毒素结合蛋白质配基,为其在内毒素致病作用机理及在内毒素血症防治研究中的应用奠定基础.方法:以内毒素为靶分子从随机七肽噬菌体展示库中筛选内毒素的高亲和力噬菌体配体,通过ELISA鉴定,DNA测序及相关软件分析.结果:所筛选的亲和力最高的噬菌体的ELISA检测值A405nm可达1.965通过比较亲和性噬菌体外源插入肽的DNA序列,认为FHENWPS肽段中包含有与内毒素分子发生亲和结合的一个共有序列.该序列展示肽的等电点为5.36,具有双嗜性,这有利于肽与LPS分子表面的位点相互作用从而产生亲和吸附.结论:运用亲和筛选方法从肽库中筛选内毒素结合蛋白质配基是可行的.     

噬菌体抗体库技术在肿瘤抗体药物研究中的进展及应用

目的:综述噬菌体抗体库技术的研究进展,介绍该技术的原理,构建,筛选和应用,为抗肿瘤抗体药物研发提供参考。方法:采用文献综述的方法,筛选近5年来噬菌体抗体库技术试验论文,对噬菌体抗体库技术的原理,构建,筛选和应用进行总结。结果:噬菌体抗体库主要分为免疫抗体库和非免疫抗体库两大类;噬菌体抗体库筛选技术包括亲和筛选、细胞筛选和生物体内筛选三种;噬菌体抗体库技术主要应用于肿瘤标志物的识别和肿瘤诊断,抗肿瘤抗体药物的筛选和制备。结论:噬菌体抗体库技术方便、快速、高效,可以在体外环境下培养,这些特点决定了其在肿瘤标志物的发现和肿瘤抗体药物研发中的广泛应用。目前噬菌体抗体库技术还存在一定缺陷,但技术的不断发展和革新必然使噬菌体抗体库技术成为研制抗体药物的新思路,极大促进了肿瘤抗体药物的研发。     

噬菌体肽库技术

80年代中期,George P.Smith在前人对丝状噬菌体分子生物学研究的基础上首先提出了噬菌体展示技术(Phage Display),其核心就是将蛋白质分子的表型和基因型巧妙地结合于丝状噬菌体这样一个便于对其进行一系列生化和遗传操作的载体上,从而大大简化了蛋白质分子表达库的筛选和鉴定。基于这一优点,噬菌     

噬菌体展示技术及其应用的研究进展

噬菌体展示技术因其高效、实用、便捷的优势现已广泛应用于抗原表位分析、抗体制备、药物筛选、疫苗研制以及免疫学疾病诊断、治疗等多方面的科学研究领域。现将近年来PDT的发展现状及其在生物科学领域中的应用进展综述如下。     

针对CD33分子的抗体偶联药物的研究进展

单克隆抗体因具有分子量小、毒副作用低、靶向性好等优点,近年来已成为肿瘤治疗用药的主要方式。CD33分子是免疫球蛋白超家族成员,同时也是唾液酸依赖的免疫球蛋白样凝集素家族的成员,在免疫调节过程中具有重要作用。CD33分子特异表达于白血病细胞表面而在造血干细胞中不表达,因而成为白血病免疫治疗的理想靶点。以CD33为靶点的抗体药物主要有CMA676、HUMl95、AVE9633、WM537LHIM3-4等,目前大多处于临床试验阶段。该实验室也在进行抗CD33全人源抗体的研究,利用噬菌体展示技术筛选与CD33胞外区特异性结合的单链抗体,并构建免疫毒素和抗体偶联药物以研究其体内外抗肿瘤作用。该文针对CD33分子及其抗体偶联药物的现状及趋势作一综述。     

噬菌体展示技术体内筛选的研究进展

噬菌体展示技术体内筛选是直接将噬菌体肽库注射到动物体内,筛选与活体内某些器官或组织有特异结合活性的小肽。噬菌体展示技术的体内筛选在血管靶向肽的筛选、肿瘤组织靶向肽的筛选、免疫反应的研究和相关疾病治疗、监测方面都有广泛的应用前景。     

利用噬菌体表面展示技术筛选转铁蛋白黏附肽的研究

为了筛选转铁蛋白黏附肽,应用噬菌体表面展示技术经过三轮生物淘选,成功地从随机七肽库中得到黏附转铁蛋白的重组噬菌体克隆,经过相对亲和力常数测定和DNA测序得到4个转铁蛋白黏附肽的序列。实验中以回收率和选择比为操作参数,对淘选进行了优化,并发展了一种基于噬菌体滴度的相对亲和力常数测定方法。转铁蛋白受体是一种有效的肿瘤标记物,利用转铁蛋白为载体可以实现药物靶向运输,因此转铁蛋白黏附肽将是重组蛋白质药物连接转铁蛋白的有用标签。     

Phage display: Concept,innovations, applications and future

Phage display is the technology that allows expression of exogenous (poly)peptides on the surface of phage particles. The concept is simple in principle: a library of phage particles expressing a wide diversity of peptides is used to select those that bind the desired target. The filamentous phage M13 is the most commonly used vector to create random peptide display libraries. Several methods including recombinant techniques have been developed to increase the diversity of the library. On the other extreme, libraries with various biases can be created for specific purposes. For instance, when the sequence of the peptide that binds the target is known, its affinity and selectivity can be increased by screening libraries created with limited mutagenesis of the peptide. Phage libraries are screened for binding to synthetic or native targets. The initial screening of library by basic biopanning has been extended to column chromatography including negative screening and competition between selected phage clones to identify high affinity ligands with greater target specificity. The rapid isolation of specific ligands by phage display is advantageous in many applications including selection of inhibitors for the active and allosteric sites of the enzymes, receptor agonists and antagonists, and G-protein binding modulatory peptides. Phage display has been used in epitope mapping and analysis of protein-protein interactions. The specific ligands isolated from phage libraries can be used in therapeutic target validation, drug design and vaccine development. Phage display can also be used in conjunction with other methods. The past innovations and those to come promise a bright future for this field.     

Panning and identification of a colon tumor binding peptide from a phage display peptide library

Tumor-targeting therapy can be an efficacious way to cure a malignant tumor in clinical trials. Phage display is a molecular diversity technology that allows the presentation of a large number of peptides or proteins on the surface of filamentous phage for various applications. In this study, we report on using phage display to generate peptide libraries that bind to colon cancer tissues. To accomplish this, we developed a screening protocol that contained 3 rounds of in vitro positive panning on colon cancer cells (SW480) and 2 rounds of subtractive screening in vitro on normal human intestinal epithelial cells with a phage display-7 peptide library. After several rounds of panning, both phage titer and recovery efficiency were significantly improved. Through a cell-based enzyme-linked immunosorbent assay, immunofluorescence, in vivo binding assay, immunocytochemical staining, and immunohistochemical staining, peptide CP15 (VHLGYAT) was demonstrated to be the most effective peptide in targeting tumor cells (SW480 and HT29 cells) and tumor tissues but not the normal human intestinal epithelial cells and control colon tissue. These studies suggest that peptide CP15 may be a promising lead candidate in the development of a useful colon tumor diagnostic and targeted drug delivery agent.     

噬菌体展示技术发展

噬菌体表面展示技术是一种将外源蛋白或抗体可变区与噬菌体表面特定蛋白质融合并展示于其表面,构建蛋白质或抗体库,并从中筛选特异蛋白质或抗体的基因工程技术。随着该项技术的不断完善和发展,噬菌体展示技术已被广泛应用于生命科学研究的不同领域,并显示了良好的应用前景。     

噬菌体展示技术发展

噬菌体表面展示技术是一种将外源蛋白或抗体可变区与噬菌体表面特定蛋白质融合并展示于其表面,构建蛋白质或抗体库,并从中筛选特异蛋白质或抗体的基因工程技术。随着该项技术的不断完善和发展,噬菌体展示技术已被广泛应用于生命科学研究的不同领域,并显示了良好的应用前景。     

Screening of tumor-targeting peptides for diagnosis and therapy through phage display technology

Phage display technology was introduced by G. Smith in 1985, which is highly effective in the selection of affinity peptides from a library containing billions of display peptides. The obtained peptides show potential efficacy in the development of further clinical applications, especially in tumor treatment. In this review, the basic principles, limits, developments of phage display technology and peptide libraries are introduced. Following that, the amino acid sequence of tumor target peptides for hematological and other systems are discussed. Finally, the application of target peptides in the design of imaging probes and the development of target peptide drugs for diagnosis and therapy are noted.     

Phage display screening of therapeutic peptide for cancer targeting and therapy

Recently, phage display technology has been announced as the recipient of Nobel Prize in Chemistry 2018. Phage display technique allows high affinity target-binding peptides to be selected from a complex mixture pool of billions of displayed peptides on phage in a combinatorial library and could be further enriched through the biopanning process; proving to be a powerful technique in the screening of peptide with high affinity and selectivity. In this review, we will first discuss the modifications in phage display techniques used to isolate various cancer-specific ligands by in situ, in vitro, in vivo, and ex vivo screening methods. We will then discuss prominent examples of solid tumor targeting-peptides; namely peptide targeting tumor vasculature, tumor microenvironment (TME) and overexpressed receptors on cancer cells identified through phage display screening. We will also discuss the current challenges and future outlook for targeting peptidebased therapeutics in the clinics.     

噬菌体呈现肽库技术的应用

噬菌体呈现肽库是噬菌体显示技术的一个非常重要的分支。自问世以来,随着分子生物学技术的飞速发展,它已被广泛应用于免疫学、分子生物学、药理学、疫苗学等生命科学领域。简要概述了这一技术的应用。