Antigen-specific B cell analysis
Understanding antigen-specific B cell responses and the subsequent formation of antibody-secreting cells (plasmablasts and plasma cells) is important when your therapy aims to induce antigen-specific antibodies, i.e. in vaccination studies. On the other hand, insight into B cell activation is also desirable when your treatment intends to avert antibody formation, i.e. when preventing alloimmunization, graft vs host-disease or inhibitory antibody formation against biologics.
Treatments aimed at lowering the presence of specific antibodies that contribute to pathology like autoimmune diseases, also require thorough understanding of B cell activation.
What can Sanquin do for your B cell projects?
We offer several assays to analyze the presence and characteristics of total and antigen-specific B cells in whole blood and PBMCs, as well as assays to unravel the specific antibody repertoire in longitudinal analyses of cells before and during treatment. These assays include B cell activation in whole blood, antigen-specific lymphocyte proliferation assays, ELISAs for measuring the presence of antigen-specific antibodies and isotype, and isolation of B cells using cell-sorting by flow cytometry followed by ex vivo experiments and/or B cell receptor sequencing and cloning.
Additionally, as CD4+ T cells are involved in fine-tuning B cell immune responses, it is also informative to investigate which specific CD4+ T cell populations correlate with the efficacy of an induced B cell response. These CD4+ T cells can be analyzed through extended ex vivo immunophenotyping before and during treatment, both in general or in an antigen-specific manner.
In B cell assays customized for your antigen of choice we can:
- Assess antigen-specific and immunoglobulin subclass-specific B cell frequencies and B cell maturation.
- Analyze titer, subclass specificity, affinity/avidity and specificity of target binding of specific antibodies.
- Analyze B cell antigen internalization, intracellular signaling and/or intracellular cytokine production upon antigen-specific or polyclonal B cell activation.
- Determine the effect of (immunomodulatory) compounds on B cell activation in whole blood in clinical trial settings.
- Express recombinant antibodies by cloning B cell receptors from antigen-specific B cells.
References
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Development of a SARS-CoV-2 Total Antibody Assay and the Dynamics of Antibody Response over Time in Hospitalized and Nonhospitalized Patients with COVID-19
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J Immunol 2020;205:3491-3499. doi: 10.4049/jimmunol.2000767.
Decoding the Human Immunoglobulin G-Glycan Repertoire Reveals a Spectrum of Fc-Receptor- and Complement-Mediated-Effector Activities
Dekkers G, Treffers L, Plomp R, Bentlage AEH, de Boer M, Koeleman CAM, Lissenberg-Thunnissen SN, Visser R, Brouwer M, Mok JY, Matlung H, van den Berg TK, van Esch WJE, Kuijpers TW, Wouters D, Rispens T, Wuhrer M, Vidarsson G.
Front Immunol 2017;8:877. doi: 10.3389/fimmu.2017.00877.
IgG subclasses and allotypes: from structure to effector functions
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