Isolation of phage-display library-derived scFv antibody specific to<i>Listeria monocytogenes</i>by a novel immobilized method

Nguyen, Xuan-Nhi; Trinh, Tuan-Dat; Vu, Tuyen; Le, Quang Huan; To, Kim Anh

doi:10.1111/jam.13648

Cited by 7 publications

(5 citation statements)

References 29 publications

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“…Although the metabolic function of PDC is well defined, this protein may also have a relation to pathogenicity. In Mycobacterium tuberculosis , PDC-E2 was found to induce a strong cellular response in an immunocompetent infected population, also contributing to the bacterial resistance against host reactive nitrogen intermediates 64 . In Mycobacterium bovis , PDC-E1β was identified as an immunodominant antigen in infected cattle, allowing the development of an indirect ELISA diagnostic test with better performance than commercial assays 65 .…”

Section: Discussionmentioning

confidence: 99%

Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

Moreira

Köllner

Helmsing

et al. 2020

Sci Rep

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The genus Listeria comprises ubiquitous bacteria, commonly present in foods and food production facilities. In this study, three different phage display technologies were employed to discover targets, and to generate and characterize novel antibodies against Listeria: antibody display for biomarker discovery and antibody generation; ORFeome display for target identification; and single-gene display for epitope characterization. With this approach, pyruvate dehydrogenase complex—enzyme 2 (PDC-E2) was defined as a new detection target for Listeria, as confirmed by immunomagnetic separation-mass spectrometry (IMS-MS). Immunoblot and fluorescence microscopy showed that this protein is accessible on the bacterial cell surface of living cells. Recombinant PDC-E2 was produced in E. coli and used to generate 16 additional antibodies. The resulting set of 20 monoclonal scFv-Fc was tested in indirect ELISA against 17 Listeria and 16 non-Listeria species. Two of them provided 100% sensitivity (CI 82.35–100.0%) and specificity (CI 78.20–100.0%), confirming PDC-E2 as a suitable target for the detection of Listeria. The binding region of 18 of these antibodies was analyzed, revealing that ≈ 90% (16/18) bind to the lipoyl domains (LD) of the target. The novel target PDC-E2 and highly specific antibodies against it offer new opportunities to improve the detection of Listeria.

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Section: Discussionmentioning

confidence: 99%

Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

Moreira

Köllner

Helmsing

et al. 2020

Sci Rep

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“…To select specific antibodies against a given antigen in its biological context, several studies have performed Phage Display selection using whole cells, isolated or in tissue conformation. This type of selection is often adopted against membrane antigens, whose native conformation is crucial to success in obtaining antibodies that bind to the target in vivo [ 68 , 69 , 93 ].…”

Section: Biopanning Selection For Retrieval Of Specific Antibodiesmentioning

confidence: 99%

Progress on Phage Display Technology: Tailoring Antibodies for Cancer Immunotherapy

França,

Studart,

Bezerra

et al. 2023

Viruses

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The search for innovative anti-cancer drugs remains a challenge. Over the past three decades, antibodies have emerged as an essential asset in successful cancer therapy. The major obstacle in developing anti-cancer antibodies is the need for non-immunogenic antibodies against human antigens. This unique requirement highlights a disadvantage to using traditional hybridoma technology and thus demands alternative approaches, such as humanizing murine monoclonal antibodies. To overcome these hurdles, human monoclonal antibodies can be obtained directly from Phage Display libraries, a groundbreaking tool for antibody selection. These libraries consist of genetically engineered viruses, or phages, which can exhibit antibody fragments, such as scFv or Fab on their capsid. This innovation allows the in vitro selection of novel molecules directed towards cancer antigens. As foreseen when Phage Display was first described, nowadays, several Phage Display-derived antibodies have entered clinical settings or are undergoing clinical evaluation. This comprehensive review unveils the remarkable progress in this field and the possibilities of using clever strategies for phage selection and tailoring the refinement of antibodies aimed at increasingly specific targets. Moreover, the use of selected antibodies in cutting-edge formats is discussed, such as CAR (chimeric antigen receptor) in CAR T-cell therapy or ADC (antibody drug conjugate), amplifying the spectrum of potential therapeutic avenues.

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“…Besides ex vivo hybridoma cell technology and in vivo production in mammalians, antibodies can be manufactured alternatively through exploiting phage-display banks [67]. In fact, phage-display technology was used to produce single-chain fragment (scFv) binders from different clones to bind a wide range of Listeria monocytogenes serotypes and strains where "traditional" methods failed in many attempts [68]. Finally, specific non-antibody binders, such as aptamers and molecular imprinted polymers (MIPs), as alternatives for antibodies are attracting increasing attention as well.…”

Section: Antigens and Antibodies As Potential Analytical Toolsmentioning

confidence: 99%

Modernization of Control of Pathogenic Micro-Organisms in the Food-Chain Requires a Durable Role for Immunoaffinity-Based Detection Methodology—A Review

Bergwerff

Debast

2021

Foods

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Food microbiology is deluged by a vastly growing plethora of analytical methods. This review endeavors to color the context into which methodology has to fit and underlines the importance of sampling and sample treatment. The context is that the highest risk of food contamination is through the animal and human fecal route with a majority of foodborne infections originating from sources in mass and domestic kitchens at the end of the food-chain. Containment requires easy-to-use, failsafe, single-use tests giving an overall risk score in situ. Conversely, progressive food-safety systems are relying increasingly on early assessment of batches and groups involving risk-based sampling, monitoring environment and herd/flock health status, and (historic) food-chain information. Accordingly, responsible field laboratories prefer specificity, multi-analyte, and high-throughput procedures. Under certain etiological and epidemiological circumstances, indirect antigen immunoaffinity assays outperform the diagnostic sensitivity and diagnostic specificity of e.g., nucleic acid sequence-based assays. The current bulk of testing involves therefore ante- and post-mortem probing of humoral response to several pathogens. In this review, the inclusion of immunoglobulins against additional invasive micro-organisms indicating the level of hygiene and ergo public health risks in tests is advocated. Immunomagnetic separation, immunochromatography, immunosensor, microsphere array, lab-on-a-chip/disc platforms increasingly in combination with nanotechnologies, are discussed. The heuristic development of portable and ambulant microfluidic devices is intriguing and promising. Tant pis, many new platforms seem unattainable as the industry standard. Comparability of results with those of reference methods hinders the implementation of new technologies. Whatever the scientific and technological excellence and incentives, the decision-maker determines this implementation after weighing mainly costs and business risks.

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Isolation of phage-display library-derived scFv antibody specific toListeria monocytogenesby a novel immobilized method

Cited by 7 publications

References 29 publications

Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

Progress on Phage Display Technology: Tailoring Antibodies for Cancer Immunotherapy

Modernization of Control of Pathogenic Micro-Organisms in the Food-Chain Requires a Durable Role for Immunoaffinity-Based Detection Methodology—A Review

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