Inverted B Cell ImmunoSpot Kits
Why use the Inverted B cell ImmunoSpot® assay?
Are you struggling to detect antigen-reactive antibody-secreting cells (ASC) in ImmunoSpot® assays despite leveraging the affinity capture coating approach to achieve high density coating (Publication) Is the coating concentration required for achieving maximal sensitivity of antigen-reactive ASC prohibitively high? If so, have you instead considered “inverting” the assay and using the affinity-tagged antigen itself as the detection probe? Moreover, did you know that inverted ImmunoSpot® assays permit assessment of ASC cross-reactivity and/or affinity at single-cell resolution?
Antigen Coating – For some antigens, particularly those with a small molecular weight (>25 kDa), achieving high density coating can be challenging and, in some instances, may require prohibitively high concentrations. Owing to the high capacity of PVDF membranes to bind immunoglobulin (Ig) and the highly efficient capture of secreted antibody achieved using Ig class (or subclass)-specific capture reagents, the inverted assay approach may enable detection of antigen-specific ASC that otherwise would be ‘undetectable.’
Detection of rare ASC producing Ig or IgG subclass – Instead of capturing all ASC-derived footprints using light chain-specific capture reagents, the inverted assay approach enables selective capture of only the Ig class or IgG subclass of interest. Moreover, selective capture of a particular Ig class or IgG subclass circumvents the challenges associated with detecting rare antigen-specific reactive ASC in competition with the other more abundant ASC populations.
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