High-Resolution Epitope Mapping and Affinity Binding Analysis Comparing a New Anti-Human LAG3 Rabbit Antibody Clone to the Commonly Used Mouse 17B4 Clone

Warren, P. Daniel; Dodson, Mark S.; Smith, Micah J.; Landowski, Terry H.; Palting, John D.

doi:10.3390/antib11040060

Cited by 3 publications

(3 citation statements)

References 35 publications

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“…SPR epitope binding analysis was performed as previously described 17 . Briefly, SPR kinetic analyses of peptide binding of the Ber‐H2 clone were performed on a BIAcore T200 using Series S CM5 sensor chips.…”

Section: Methodsmentioning

confidence: 99%

“…SPR epitope binding analysis was performed as previously described. 17 Briefly, SPR kinetic analyses of peptide binding of the Ber-H2 clone were performed on a BIAcore T200 using Series S CM5 sensor chips. The mouse antibody capture surface was prepared with 50 ng/mL goat anti-species IgG Fc-specific in 10 mM acetate pH 5.…”

Section: Spr Epitope Binding Kinetics Assaymentioning

confidence: 99%

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Anti‐CD30 (Ber‐H2) epitope requires structural elements as shown by mass spectroscopy and dual‐site associated kinetics

Warren

Smith

2023

J of Molecular Recognition

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The Ber‐H2 mouse monoclonal antibody has been in use for 35 years for detecting the CD‐30 biomarker in a variety of lymphomas. Despite the wide use of this clone, we have not been successful in applying synthetic peptides derived from the published epitope sequence and affinity data toward the development of a new Ber‐H2‐based in vitro diagnostic reagent assay. We found that synthetic peptides based on the published epitope sequence do not function to inhibit antibody‐binding activity, thus indicating that the sequence is not the full epitope recognized by Ber‐H2. In this report, we used mass spectroscopic analysis of proteolyzed CD30 fragments capable of binding Ber‐H2 to identify additional regions within the epitope that participate in binding. Using surface plasmon resonance binding kinetic analyses and immuno‐histochemical peptide‐inhibition assays, we also demonstrate that the epitope sequence as originally reported is missing two key elements necessary for binding the Ber‐H2 antibody.

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Section: Methodsmentioning

confidence: 99%

Section: Spr Epitope Binding Kinetics Assaymentioning

confidence: 99%

Anti‐CD30 (Ber‐H2) epitope requires structural elements as shown by mass spectroscopy and dual‐site associated kinetics

Warren

Smith

2023

J of Molecular Recognition

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“…Mice and rats are widely used for immunization to generate mAbs; however, the use of rabbits for the mAb development has attracting interest because of their unique and highly distinctive antibody repertoire, which exhibits high-affinity, specificity, and diversity [ 18 ]. Indeed, previous studies have been succeeded in developing high-affinity mAbs to several antigens using rabbits [ 19 – 21 ]. These findings prompted us to establish a rabbit mAb against bovine PD-1.…”

Section: Introductionmentioning

confidence: 99%

Development of a high-affinity anti-bovine PD-1 rabbit–bovine chimeric antibody using an efficient selection and large production system

Okagawa,

Konnai,

Goto

et al. 2023

Vet Res

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Immune checkpoint molecules PD-1/PD-L1 cause T-cell exhaustion and contribute to disease progression in chronic infections of cattle. We established monoclonal antibodies (mAbs) that specifically inhibit the binding of bovine PD-1/PD-L1; however, conventional anti-PD-1 mAbs are not suitable as therapeutic agents because of their low binding affinity to antigen. In addition, their sensitivity for the detection of bovine PD-1 is low and their use for immunostaining PD-1 is limited. To address these issues, we established two anti-bovine PD-1 rabbit mAbs (1F10F1 and 4F5F2) and its chimeric form using bovine IgG1 (Boch1D10F1), which exhibit high binding affinity. One of the rabbit mAb 1D10F1 binds more strongly to bovine PD-1 compared with a conventional anti-PD-1 mAb (5D2) and exhibits marked inhibitory activity on the PD-1/PD-L1 interaction. In addition, PD-1 expression in bovine T cells could be detected with higher sensitivity by flow cytometry using 1D10F1. Furthermore, we established higher-producing cells of Boch1D10F1 and succeeded in the mass production of Boch1D10F1. Boch1D10F1 exhibited a similar binding affinity to bovine PD-1 and the inhibitory activity on PD-1/PD-L1 binding compared with 1D10F1. The immune activation by Boch1D10F1 was also confirmed by the enhancement of IFN-γ production. Finally, Boch1D10F1 was administered to bovine leukemia virus-infected cows to determine its antiviral effect. In conclusion, the high-affinity anti-PD-1 antibody developed in this study represents a powerful tool for detecting and inhibiting bovine PD-1 and is a candidate for PD-1-targeted immunotherapy in cattle.

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Lymphocyte activation gene-3 as a candidate target for combo anti-programmed death-(ligand) 1 therapy of cancer

Mortezaee

2024

Process Biochemistry

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High-Resolution Epitope Mapping and Affinity Binding Analysis Comparing a New Anti-Human LAG3 Rabbit Antibody Clone to the Commonly Used Mouse 17B4 Clone

Cited by 3 publications

References 35 publications

Anti‐CD30 (Ber‐H2) epitope requires structural elements as shown by mass spectroscopy and dual‐site associated kinetics

Anti‐CD30 (Ber‐H2) epitope requires structural elements as shown by mass spectroscopy and dual‐site associated kinetics

Development of a high-affinity anti-bovine PD-1 rabbit–bovine chimeric antibody using an efficient selection and large production system

Lymphocyte activation gene-3 as a candidate target for combo anti-programmed death-(ligand) 1 therapy of cancer

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