Binding Affinity Measurement of Antibodies from Crude Hybridoma Samples by SPR

Ndao, Dorin Mlaki; Hickman, David T.; López-Deber, María Pilar; Davranche, Aurélien; Pfeifer, Andréa; Muhs, Andreas

doi:10.21769/bioprotoc.1276

Cited by 2 publications

(2 citation statements)

References 3 publications

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“…Herein, the term crude refers to: a mixture of un-purified, cell culture media with variable specific antibody concentrations. Screening crude monoclonal antibodies (mAbs) saves time and money in comparison with first purifying a panel of mAbs and then testing them all for application in LFIA [10]. Previously, true kinetic studies have been carried out to select antibodies based on their affinities, association and dissociation rates, for application in a direct SPR biosensor [11].…”

Section: Introductionmentioning

confidence: 99%

Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow Prototypes

et al. 2018

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Lateral Flow Immunoassays (LFIAs) allow for rapid, low-cost, screening of many biomolecules such as food allergens. Despite being classified as rapid tests, many LFIAs take 10–20 min to complete. For a really high-speed LFIA, it is necessary to assess antibody association kinetics. By using a label-free optical technique such as Surface Plasmon Resonance (SPR), it is possible to screen crude monoclonal antibody (mAb) preparations for their association rates against a target. Herein, we describe an SPR-based method for screening and selecting crude anti-hazelnut antibodies based on their relative association rates, cross reactivity and sandwich pairing capabilities, for subsequent application in a rapid ligand binding assay. Thanks to the SPR selection process, only the fast mAb (F-50-6B12) and the slow (S-50-5H9) mAb needed purification for labelling with carbon nanoparticles to exploit high-speed LFIA prototypes. The kinetics observed in SPR were reflected in LFIA, with the test line appearing within 30 s, almost two times faster when F-50-6B12 was used, compared with S-50-5H9. Additionally, the LFIAs have demonstrated their future applicability to real life samples by detecting hazelnut in the sub-ppm range in a cookie matrix. Finally, these LFIAs not only provide a qualitative result when read visually, but also generate semi-quantitative data when exploiting freely downloadable smartphone apps.

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Section: Introductionmentioning

confidence: 99%

Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow Prototypes

et al. 2018

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“…After the Ab is produced, as described before, surface plasmon resonance (SPR), equilibrium dialysis, ELISA, or many other methods are widely used to indicate its affinity (termed as binding strength or binding constant) to the Ag, and demonstrate its characteristics and binding to the target drug in real-time, and in a label-free manner, using a refractive index change at a metal surface [ 53 , 54 ]. There is also the possibility of using ELISA to verify that the Ab meets the need with the target drug (i.e., sensitivity, specificity).…”

Section: Performances Of An Antibodymentioning

confidence: 99%

Monoclonal Antibodies Application in Lateral Flow Immunochromatographic Assays for Drugs of Abuse Detection

et al. 2021

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Lateral flow assays (lateral flow immunoassays and nucleic acid lateral flow assays) have experienced a great boom in a wide variety of early diagnostic and screening applications. As opposed to conventional examinations (High Performance Liquid Chromatography, Polymerase Chain Reaction, Gas chromatography-Mass Spectrometry, etc.), they obtain the results of a sample’s analysis within a short period. In resource-limited areas, these tests must be simple, reliable, and inexpensive. In this review, we outline the production process of antibodies against drugs of abuse (such as heroin, amphetamine, benzodiazepines, cannabis, etc.), used in lateral flow immunoassays as revelation or detection molecules, with a focus on the components, the principles, the formats, and the mechanisms of reaction of these assays. Further, we report the monoclonal antibody advantages over the polyclonal ones used against drugs of abuse. The perspective on aptamer use for lateral flow assay development was also discussed as a possible alternative to antibodies in view of improving the limit of detection, sensitivity, and specificity of lateral flow assays.

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Binding Affinity Measurement of Antibodies from Crude Hybridoma Samples by SPR

Cited by 2 publications

References 3 publications

Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow Prototypes

Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow Prototypes

Monoclonal Antibodies Application in Lateral Flow Immunochromatographic Assays for Drugs of Abuse Detection

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