“Cleavable” hapten–biotin conjugates: Preparation and use for the generation of anti-steroid single-domain antibody fragments

Kobayashi, N.; Oyama, Hiroyuki; Nakano, Masanori; Kanda, Tatsuaki; Banzono, Erika; Kato, Yoshinori; Karibe, Tsuyoshi; Nishio, Takehiko; Goto, Junichi

doi:10.1016/j.ab.2009.01.004

Cited by 12 publications

(15 citation statements)

References 46 publications

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“…We have already undertaken modification of the present CAP into more efficient and feasible procedures. To surely recover scFv mutants bound on the antigen-immobilized solid phases with extremely high affinity, we might employ the dissociation-independent methods such that we previously reported 14 , 42 , 43 .…”

Section: Discussionmentioning

confidence: 99%

Clonal array profiling of scFv-displaying phages for high-throughput discovery of affinity-matured antibody mutants

Kiguchi

Oyama

Morita

et al. 2020

Sci Rep

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"Antibody-breeding" approach potentially generates therapeutic/diagnostic antibody mutants with greater performance than native antibodies. Therein, antibody fragments (e.g., single-chain Fv fragments; scFvs) with a variety of mutations are displayed on bacteriophage to generate diverse phage-antibody libraries. Rare clones with improved functions are then selected via panning against immobilized or tagged target antigens. However, this selection process often ended unsuccessful, mainly due to the biased propagation of phage-antibody clones and the competition with a large excess of undesirable clones with weaker affinities. To break radically from such panning-inherent problems, we developed a novel method, clonal array profiling of scFv-displaying phages (CAP), in which colonies of the initial bacterial libraries are examined one-by-one in microwells. Progenies of scFv-displaying phages generated are, if show sufficient affinity to target antigen, captured in the microwell via pre-coated antigen and detected using a luciferase-fused anti-phage scFv. The advantage of CAP was evidenced by its application with a small error-prone-PCR-based library (~ 10 5 colonies) of anti-cortisol scFvs. Only two operations, each surveying only ~ 3% of the library (9,400 colonies), provided five mutants showing 32–63-fold improved K a values (> 10 10 M −1 ), compared with the wild-type scFv ( K a = 3.8 × 10 8 M −1 ), none of which could be recovered via conventional panning procedures operated for the entire library.

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Section: Discussionmentioning

confidence: 99%

Clonal array profiling of scFv-displaying phages for high-throughput discovery of affinity-matured antibody mutants

Kiguchi

Oyama

Morita

et al. 2020

Sci Rep

Self Cite

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“…Nevertheless, there are several examples showing that VHHs can be exploited for detecting haptens as different as herbicides, caffeine, mycotoxins, trinitrotoluene, steroids and therapeutic drugs [113-120] for this antibody class can interact with its targets adopting different binding patterns [121]. For instance, the CDR1 loop provides a strong interaction for the azo-dye Reactive Red 6 [122], CDR2 and a framework residue contribute to the binding of Reactive Red 1 [123], whereas a non-conventional, substrate-dependent dimerization mechanism is involved in the binding of caffeine and its metabolites [124].…”

Section: Introductionmentioning

confidence: 99%

Biotechnological applications of recombinant single-domain antibody fragments

Marco

2011

Microb Cell Fact

147

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BackgroundSingle-domain antibody fragments possess structural features, such as a small dimension, an elevated stability, and the singularity of recognizing epitopes non-accessible for conventional antibodies that make them interesting for several research and biotechnological applications.ResultsThe discovery of the single-domain antibody's potentials has stimulated their use in an increasing variety of fields. The rapid accumulation of articles describing new applications and further developments of established approaches has made it, therefore, necessary to update the previous reviews with a new and more complete summary of the topic.ConclusionsBeside the necessary task of updating, this work analyses in detail some applicative aspects of the single-domain antibodies that have been overseen in the past, such as their efficacy in affinity chromatography, as co-crystallization chaperones, protein aggregation controllers, enzyme activity tuners, and the specificities of the unconventional single-domain fragments.

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“…However, the display of antigenic epitopes in a properly folded and functioning conformation that could elicit a strong immune response is a significant issue which deserves further attention in the development of phage display vaccines. Sometimes, rather than constructing a transcriptional fusion to a coat protein, phage disply vaccines may be created by conjugating the antigenic substance to the phage surface via artificial linkers [ 87 ]. The exploitation of this strategy for the construction of phage display vaccines may diversify the range of antigens that can be displayed on the surface of immunogenic phage particles.…”

Section: Phage Display Vaccines: Bacterial Viruses En Route To Antigementioning

confidence: 99%

Phage display as a promising approach for vaccine development

et al. 2016

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Bacteriophages are specific antagonists to bacterial hosts. These viral entities have attracted growing interest as optimal vaccine delivery vehicles. Phages are well-matched for vaccine design due to being highly stable under harsh environmental conditions, simple and inexpensive large scale production, and potent adjuvant capacities. Phage vaccines have efficient immunostimulatory effects and present a high safety profile because these viruses have made a constant relationship with the mammalian body during a long-standing evolutionary period. The birth of phage display technology has been a turning point in the development of phage-based vaccines. Phage display vaccines are made by expressing multiple copies of an antigen on the surface of immunogenic phage particles, thereby eliciting a powerful and effective immune response. Also, the ability to produce combinatorial peptide libraries with a highly diverse pool of randomized ligands has transformed phage display into a straightforward, versatile and high throughput screening methodology for the identification of potential vaccine candidates against different diseases in particular microbial infections. These libraries can be conveniently screened through an affinity selection-based strategy called biopanning against a wide variety of targets for the selection of mimotopes with high antigenicity and immunogenicity. Also, they can be panned against the antiserum of convalescent individuals to recognize novel peptidomimetics of pathogen-related epitopes. Phage display has represented enormous promise for finding new strategies of vaccine discovery and production and current breakthroughs promise a brilliant future for the development of different phage-based vaccine platforms.

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“Cleavable” hapten–biotin conjugates: Preparation and use for the generation of anti-steroid single-domain antibody fragments

Cited by 12 publications

References 46 publications

Clonal array profiling of scFv-displaying phages for high-throughput discovery of affinity-matured antibody mutants

Clonal array profiling of scFv-displaying phages for high-throughput discovery of affinity-matured antibody mutants

Biotechnological applications of recombinant single-domain antibody fragments

Phage display as a promising approach for vaccine development

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