Production and Characterization of Murine Single Chain Fv Antibodies to Aflatoxin B 1 Derived From a Pre-immunized Antibody Phage Display Library System

Daly, Stephen J.; Dillon, Paul P.; Manning, Bernadette M.; Dunne, Lynsey; Killard, Anthony J.; O’Kennedy, Richard

doi:10.1080/0954010021000096373

Cited by 42 publications

(27 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Specific scFv have been developed for a number of mycotoxins including zearalenone (Chang, Choi, & Chun, 2008;Wang, Du, Lin, Huang, & Wang, 2008;Yuan et al, 1997), aflatoxin B 1 (Daly et al, 2002;Moghaddam, Løbersli, Gebhardt, Braunagel, & Marvik, 2001) and fumonisin B 1 (Lauer, Ottleben, Jacobsen, & Reinard, 2005;Min et al, 2010). There are also two reports of scFv specific for DON (Choi et al, 2004;Wang et al, 2007) and a report of a single-domain antibody fragment with an affinity to a related congener, 15-acetyl-deoxynivalenol (Doyle et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Production and characterization of a single chain variable fragment (scFv) against the mycotoxin deoxynivalenol

Maragos

Chen

2012

Food and Agricultural Immunology

View full text Add to dashboard Cite

Deoxynivalenol (DON) is a mycotoxin produced by certain fungi that infest cereal grains. A hybridoma cell line producing a monoclonal antibody (Mab) was used as the starting point in the development of a recombinant single chain variable fragment antibody (scFv) recognising DON. The scFv and Mab were characterised using two immunoassay formats: competitive direct enzyme-linked immunosorbent assay (CD-ELISA) and biolayer interferometry (BLI). Using CD-ELISA the IC 50 s for DON were 36.1 and 13.8 ng/ml for assays based on the scFv and Mab, respectively. The cross-reactivity to DON analogs was very similar for the scFv and the Mab. The real-time binding of the antibodies to an immobilised DON-protein conjugate was also monitored. In competitive BLI assays the IC 50 s using the scFv and Mab were 68.3 and 15.8 ng/ml, respectively. The results suggest that sensitivity of assays, but not selectivity, was affected by removal of the constant regions of the Mab.

show abstract

Section: Introductionmentioning

confidence: 99%

Production and characterization of a single chain variable fragment (scFv) against the mycotoxin deoxynivalenol

Maragos

Chen

2012

Food and Agricultural Immunology

View full text Add to dashboard Cite

show abstract

“…Since obtaining specific antibody to free hapten is often very difficult with either naïve libraries or libraries derived from immunized animals, many strategies have been developed to increase the chance of selecting specific antibodies from a phage display antibody library. These include modification of the elution step by competing with free hapten [18], alternative hapten-carrier conjugates [6], selection from a focused library that favors binding haptens (cavity library) [19], or selection from an immunized library [20]. However, in this study, we were able to isolate specific antibody by using a simple bio-panning procedure.…”

Section: Discussionmentioning

confidence: 94%

“…However, they are not as abundant and toxic as AFB 1 [5]. Aflatoxin M 1 is a metabolite of AFB 1 , which can be found in milk of animals that consume aflatoxincontaminated feeds [6]. The structures of various aflatoxins are illustrated in Fig.…”

Section: Introductionmentioning

confidence: 99%

One-Step Detection of Aflatoxin-B1 Using scFv-Alkaline Phosphatase-Fusion Selected from Human Phage Display Antibody Library

et al. 2011

View full text Add to dashboard Cite

A unique human phage display library was used to successfully generate a scFv to the highly carcinogenic toxin aflatoxin B1. Such an antibody has major potential applications in therapy and diagnostics. To further exploit its analytical capacity, the scFv was genetically fused to alkaline phosphatase, thereby generating a novel and highly sensitive self-indicating reagent. The performance of this reagent was further characterized, demonstrating its efficacy. The sensitivity of scFv-AP fusion was three-fold better than that of the scFv form. The ability of this human library to generate antibodies to a small hapten was clearly demonstrated and this is linked to its intrinsic diversity, which exceeds many existing conventional human libraries. Our results indicate that demography may influence the diversity of the repertoire of the library in terms of its capacity to generate antibodies to specific targets. Equally, the approach demonstrated should also be applicable for other haptens and larger antigens.

show abstract

“…[113,127] Phagen können bei der Entwicklung von spezifischen SPR-Sensoren in zweierlei Weise genutzt werden: [110,114,113,[127][128][129] durch Verankern des zielspezifischen Phagen auf einem Sensorchip [110,127] sowie bei der Selektion eines zielspezifischen Proteins mithilfe eines Phagendisplays und anschließendem Verankern dieses Proteins auf dem Sensorchip. [113,114] Durch Einführung eines zielspezifischen Phagen in einen SPR-Sensor konnte dessen Leistungsfähigkeit merklich gesteigert werden. Beispielsweise wurde in einem ZweikanalExperiment ein b-gal-spezifischer "Landscape"-Phage, der mithilfe eines Phagendisplays identifiziert worden war (Abbildung 3), durch Physisorption auf der Goldoberfläche in einem Kanal eines SPREETA-SPR-Sensorchips verankert.…”

Section: Oberflächenplasmonenresonanz-sensorenunclassified

“…Auf der Grundlage dieses scFv-Antikörpers und der SPR wurden Inhibitionsimmunassays entwickelt, die AFB1 empfindlich und selektiv nachweisen konnten. [113] Die vorgestellten SPR-Biosensoren arbeiten viel schneller und sind viel einfacher zu handhaben als herkömmliche ELISAVerfahren.…”

Section: Oberflächenplasmonenresonanz-sensorenunclassified

Die Anwendung von Viren in Chemo‐ und Biosensoren

2009

View full text Add to dashboard Cite

Viren haben sich in jüngster Zeit als einzigartige Hilfsmittel für den empfindlichen und selektiven Nachweis von Analyten wie Explosivstoffen, Proteinen, Bakterien, Viren, Sporen und Toxinen erwiesen. Auf Bakterien spezialisierte Viren, die Bakteriophagen – oder kurz Phagen –, wurden dabei am intensivsten untersucht: Zielspezifische nichtlysierende Phagen (sowie von diesen präsentierte Peptide und Proteine) können mithilfe hoch entwickelter Phagendisplaytechniken identifiziert werden, und lysierende Phagen können gezielt Bakterien aufbrechen und spezifische Zellmarker wie Enzyme ausschütten, die analysiert werden können. Darüber hinaus sind Phagen chemisch wie thermisch hinreichend stabil, und sie lassen sich mit Molekülen kuppeln und mit Nanomaterialien kombinieren und auf der Oberfläche eines Signalwandlers in einem Analysesystems verankern. Im Mittelpunkt dieses Aufsatzes stehen Fortschritte bei der Verwendung von Phagen in Chemo‐ und Biosensoren durch Kombination mit etablierten analytischen Techniken. Überdies werden aktuelle Entwicklungen bei der Verwendung von Virus‐Nanomaterial‐Kompositen und anderen Viren in Sensoranwendungen vorgestellt.

show abstract

Production and Characterization of Murine Single Chain Fv Antibodies to Aflatoxin B 1 Derived From a Pre-immunized Antibody Phage Display Library System

Abstract: The production and characterization of recombinant antibodies to aflatoxin B 1 (AFB 1

Cited by 42 publications

References 19 publications

Production and characterization of a single chain variable fragment (scFv) against the mycotoxin deoxynivalenol

Production and characterization of a single chain variable fragment (scFv) against the mycotoxin deoxynivalenol

One-Step Detection of Aflatoxin-B1 Using scFv-Alkaline Phosphatase-Fusion Selected from Human Phage Display Antibody Library

Die Anwendung von Viren in Chemo‐ und Biosensoren

Contact Info

Product

Resources

About