Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance

mAbs, Journal Year: 2023, Volume and Issue: 15(1)

Published: Dec. 13, 2023

Accurate and efficient affinity measurement techniques are essential for the biophysical characterization of therapeutic monoclonal antibodies, one fastest growing drug classes. Surface plasmon resonance (SPR) is widely used determining antibody affinity, but does not perform well with extremely high (low picomolar to femtomolar range) molecules. In this study, we compare SPR-based Carterra LSA kinetic exclusion assay (KinExA) measuring affinities 48 antibodies generated against SARS-CoV-2 receptor-binding domain. These data reveal that high-affinity can be straight from selections using high-quality in vitro library platforms 54% correspondence between measured KinExA. Generally, where there was a 2-fold or greater difference KinExA, KinExA reported were tighter. We highlight differences identifying benefits pitfalls each terms dynamic range throughput. Furthermore, demonstrate first time single-point screening significantly improve throughput while maintaining strong correlation full binding curve equilibrium measurements, enabling accurate rank-ordering clones exceptionally tight properties.

Language: Английский

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Precision Targeting and Genetically Modified T Cells for Targeting Cancer Cells

International Journal of Trends in OncoScience, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 9

Published: Jan. 5, 2024

In this review cancer treatment, despite notable progress, challenges persist globally. Traditional methods like surgery, chemotherapy, and radiotherapy, while effective, often compromise patients' overall quality of life due to side effects. Immunotherapeutic strategies, especially Chimeric Antigen Receptor T cells, show promise by leveraging the immune system target tumors independently certain escape mechanisms. However, CAR-T cells' specificity surface antigens limits their applicability. Precise management demands ongoing research refine broaden these therapies. Employing CAR or T-cell receptor therapies, genetic engineering enhances antigenic specificity, optimizing immunotherapy precision. CARs, synthetic receptors engineered for tumor antigen recognition, represent a groundbreaking approach, intertwining immunotherapy, gene therapy, therapy. The human system's ability discern self from non-self-entities forms basis fostering innovative modalities that selectively cells. with FDA approval leukemia lymphoma, holds transformative potential but faces safety efficacy challenges. Advances, including mitigating cytotoxicity enhancing therapeutic efficacy, promise. Utilizing alteration, CARs have shown in treatment hematologic malignancies, particularly CD19 B cell blood cancers. Current study is investigating uses patients lymphoma myeloma.

Language: Английский

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Optical sensing and control of T cell signaling pathways

Frontiers in Physiology, Journal Year: 2024, Volume and Issue: 14

Published: Jan. 10, 2024

T cells regulate adaptive immune responses through complex signaling pathways mediated by cell receptor (TCR). The functional domains of the TCR are combined with specific antibodies for development chimeric antigen (CAR) therapy. In this review, we first overview current understanding on as well traditional methods that have been widely used study. These methods, however, still limited to investigating dynamic molecular events spatiotemporal resolutions. Therefore, genetically encoded biosensors and optogenetic tools developed study in live cells. We review these cutting-edge technologies revealed mechanisms at each stage pathways. They primarily applied signaling, they will further aid CAR activation function. offer powerful enhancing our CAR-T

Language: Английский

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A method for rapid nanobody screening with no bias of the library diversity

iScience, Journal Year: 2024, Volume and Issue: 27(2), P. 108966 - 108966

Published: Jan. 18, 2024

Nanobody, referred to the variable domain of heavy-chain-only antibodies, has several advantages such as small size and feasible

Language: Английский

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CARs derived from broadly neutralizing, human monoclonal antibodies identified by single B cell sorting target hepatitis B virus-positive cells

Frontiers in Immunology, Journal Year: 2024, Volume and Issue: 15

Published: April 22, 2024

To design new CARs targeting hepatitis B virus (HBV), we isolated human monoclonal antibodies recognizing the HBV envelope proteins from single cells of a patient with resolved infection. HBV-specific memory were by incubating peripheral blood mononuclear biotinylated surface antigen (HBsAg), followed single-cell flow cytometry-based sorting live, CD19 + IgG HBsAg cells. Amplification and sequencing immunoglobulin genes identified variable heavy light chain sequences. Corresponding chains cloned into IgG1 expression vectors expressed in mammalian Two named 4D06 4D08 found to be highly specific for HBsAg, recognized conformational linear epitope, respectively, showed broad reactivity neutralization capacity against all major genotypes. fragments 2 nd generation CAR format CD28 CD3zeta intracellular signaling domains. The constructs displayed high functional avidity when on primary T CAR-grafted proved polyfunctional regarding cytokine secretion killed HBV-positive target Interestingly, background activation 4D08-CAR instead epitope was consistently low. In preclinical model chronic infection, murine grafted activity indicated transient increase serum transaminases, lower number hepatocytes mice treated. This study demonstrates an efficient fast approach identifying pathogen-specific small donor cell numbers subsequent CARs.

Language: Английский

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