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... the introduction of phage display, other display systems have been developed. This includes systems like yeast display, bacterial cell surface display, ribosomal display, mRNA display, DNA display and mammalian cell surface display [17]. Figure 1 shows the alterna- tive display systems used for antibody presentation. Yeast display has an additional feature compared to phage display when dealing with mammalian proteins. This is due to the appli- cation of the eukaryotic machinery to assimilate the display mechanism. In this system, the antibody gene is fused to the Aga2p agglutinin subunit found on the surface of yeast cells [18,19]. In a similar fashion, bacterial cell display functions by displaying antibodies on the surface of Gram-negative or Gram-positive cells as a fusion to the flagella or outer membrane proteins ...
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... introduction of recombinant antibody technology has revolutionized and improved the way antibodies are being generated for various applications in research, diagnosis and therapy [1][2][3][4]. Antibodies have been the cornerstone for many biomedical advances in the past due to its high specificity and affinity to capture target antigens. The key characteristic of antibodies that makes it highly sought after is the defined specificity of the complemen- tarity determining regions (CDR) of the variable domains against a specific target [5]. This specificity is programmed in vivo by a series of different molecular mechanisms such as V-D-J recombination of the heavy chain, V-J recombination of the light chain and somatic hyper- mutation [6,7]. After primary immune response the V H D H J H and V L J L exons are randomly mutated, mainly in the CDRs, by somatic hypermutation leading to high affinity antibod- ies (see article of Oliver Backhaus in this book). These molecular processes have a pro- found effect on the way the genotype is delineated as the gene rearrangements will bring about multiple gene segment combinations. Additional mutagenesis is elicited through incorporation of additional nucleotides between the junctions of the V, D and J gene seg- ment of the heavy chain and V and J gene segment of the light chain. These variations at the genotypic level have a direct influence on the phenotypic variation seen in terms of target specificity and affinity of the generated antibody [8,9]. Figure 1 shows the cor- relation between the genotypic variations and the phenotypic nature of the generated antibodies. The introduction of recombinant DNA technology and display technologies has allowed recombinant antibodies to be generated at a rapid pace. This is evident with the increase of recombinant antibodies going into clinical trials in the last 3 years [10,11]. The general con- cept of display oriented techniques for antibody generation relies upon the ability to harness the natural or synthetic diversity of an antibody library [12]. As with most recombinant DNA approaches, the ability to customize or modify the genotype either at single base or amino acid level was now possible [13]. This opened many new avenues in the field of recombinant antibody technology to allow modification and customization of characteristics of the pheno- type. The advent of display technologies allowed for selective isolation of specific phenotypes with their respective genotypic information to be retrieved together [14]. This means that it was now possible to replicate the in vivo antibody generation and maturation process in vitro [13]. The impact of antibody display technologies combined with affinity maturation strate- gies in the isolation and identification of high affinity antibodies is monumental in the way antibodies are made ...
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Citations
... Biological display systems have been systematically developed for more than 30 years and have been broadly used for high-throughput ligand selection and other fields ( Figure 2) [16]. With gene recombinant technologies, the structure of natural ligands can be conveniently imitated and reshaped by incorporating functional tags, groups, and linkers. ...
Molecular imaging plays important roles in many fields, including disease diagnosis, therapeutic efficacy evaluation, intraoperative imaging guidance, drug metabolism monitoring, and patient selection for appropriate treatment. As a key component, the targeting ligand determines the specificity, affinity, and in vivo performance of molecular imaging probes. In this review, high‐throughput screening and biological display platforms for the discovery of ligands applicable to molecular imaging are briefly reviewed. Basic information on ligand development for molecular imaging is first introduced, followed by a presentation of various selection platforms and typical or iterative cases. The features, advantages, limitations, and application scope of screening and display platforms are compared and discussed. Last, a basic selection strategy and a perspective for protein‐based ligands are provided.
... The naïve human antibody libraries have been constructed using human B cells of non-immunized or healthy donors [48]. The antibodies isolated from naïve human antibody libraries may often exhibit lower affinities because the antibody repertoire does not undergo in vivo affinity maturation to produce higher-affinity antibodies [48,49]. In the present study, we assert that the library we have developed is a high-quality human antibody library with large and diverse repertoires. ...
Antibody phage display is a key technology for the discovery and development of target-specific monoclonal antibodies (mAbs) for use in research, diagnostics, and therapy. The construction of a high-quality antibody library, with larger and more diverse antibody repertoires, is essential for the successful development of phage display-derived mAbs. In this study, a large human combinatorial single-chain variable fragment library (1.5 × 1011 colonies) was constructed from Epstein–Barr virus-infected human peripheral blood mononuclear cells stimulated with a combination of two of the activators of human B cells, the Toll-like receptor 7/8 agonist R848 and interleukin-2. Next-generation sequencing analysis with approximately 1.9 × 106 and 2.7 × 106 full-length sequences of heavy chain variable (VH) and κ light chain variable (Vκ) domains, respectively, revealed that the library consists of unique VH (approximately 94%) and Vκ (approximately 91%) sequences with greater diversity than germline sequences. Lastly, multiple unique mAbs with high affinity and broad cross-species reactivity could be isolated from the library against two therapeutically relevant target antigens, validating the library quality. These findings suggest that the novel antibody library we have developed may be useful for the rapid development of target-specific phage display-derived recombinant human mAbs for use in therapeutic and diagnostic applications.
... Types of surface display systems. The three main systems are based on virus-surface, cell-surface and cell-free displays(Lim, Choong and Lim 2017). ...
Phage display technology, which is based on the presentation of peptide sequences on the surface of bacteriophage virions, was developed over 30 years ago. Improvements in phage display systems have allowed us to employ this method in numerous fields of biotechnology, as diverse as immunological and biomedical applications, the formation of novel materials and many others. The importance of phage display platforms was recognized by awarding the Nobel Prize in 2018 “for the phage display of peptides and antibodies”. In contrast to many review articles concerning specific applications of phage display systems published in recent years, we present an overview of this technology, including a comparison of various display systems, their advantages and disadvantages, and examples of applications in various fields of science, medicine, and the broad sense of biotechnology. Other peptide display technologies, which employ bacterial, yeast and mammalian cells, as well as eukaryotic viruses and cell-free systems, are also discussed. These powerful methods are still being developed and improved; thus, novel sophisticated tools based on phage display and other peptide display systems are constantly emerging, and new opportunities to solve various scientific, medical and technological problems can be expected to become available in the near future.
... B. bispezifische Antikörper, Immunzytokine oder Immuntoxine) eingesetzt. Die humanen Antikörper werden dabei aus Antikörper-Genbibliotheken mittels Phagendisplay isoliert [22,23]. Liganden, die an Zelloberflächenproteine binden, oder speziellen Trägern (Nanopartikeln) in die Tumorzelle zu transportieren. ...
Zusammenfassung
Krebsentstehung basiert auf der Anhäufung von Mutationen in Wachstumsgenen (wie z. B. Transkriptionsfaktoren, Wachstumsrezeptoren oder intrazellulären Signalmolekülen) oder in Suppressorgenen (wie z. B. p53). Während des Tumorwachstums kommt es dann zur Selektion von Zellklonen, die Mutationen in „driver genes“, die zum unkontrollierten Wachstum der Zellklone führen, enthalten. Bei allen Phasen der Tumorentwicklung (Überwachung des Tumorwachstums durch das Immunsystem, Gleichgewichtsphase, Entkommen des Tumors vor dem Immunsystem) spielen die Wechselwirkung zwischen dem Immunsystem und den Tumorzellen und die Entstehung einer chronischen Entzündung in unmittelbarer Umgebung des Tumors eine entscheidende Rolle. Die Immuntherapie ist eine Krebstherapie, die das Immunsystem aktivieren soll. Eine vielversprechende angewandte Immuntherapie basiert auf Antikörpern, die Immunzellen aktivieren, das Tumorwachstum inhibieren oder zur Eliminierung der Tumorzellen führen. Dabei werden rekombinante IgG-Antikörper oder gentechnologisch veränderte Antikörperfragmente gegen tumorassoziierte Antigene (TAA’s) einzeln oder in Kombination mit Chemo- oder Strahlentherapie eingesetzt. Vielversprechend und zugelassen sind Checkpointantikörper, welche die Blockade von zytotoxischen CD8 ⁺ -T-Zellen und CD4 ⁺ -T-Zellen durch Tumorzellen und/oder dendritische Zellen aufheben. Andere erfolgreiche Antikörperkonstrukte sind bispezifische Antikörper (binden an T‑Zelle und Tumorzelle), chimäre Antigenrezeptoren (CAR) für die T‑Zell-Therapie, Immuntoxine (Antikörper fusioniert mit einem Toxin) und Immunzytokine (Antikörper fusioniert mit einem Zytokin). Außerdem haben intrazelluläre Antikörper, die erfolgreich in Xenograft-Tumor-Mausmodellen getestet worden sind, vielversprechendes therapeutisches Potenzial.
