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10.3390/cells10081843 http://scihub22266oqcxt.onion/10.3390/cells10081843 34440612!8394687!34440612     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Cells 2021 ; 10 (8): ä Nephropedia Template TP {{tp|p=34440612|t=2021. Affinity Tag Coating Enables Reliable Detection of Antigen-Specific B Cells in Immunospot Assays |pdf=|usr=}}{{34440612}} gab.com Text Affinity Tag Coating Enables Reliable Detection of Antigen-Specific B Cells in Immunospot Assays JO: Cells http://www.kidney.de/pget.php?&tw=1&pmid=34440612 #FOAvip Assessment of humoral immunity to SARS-CoV-2 and other infectious agents is typically restricted to detecting antigen-specific antibodies in the serum. Rarely does immune monitoring entail assessment of the memory B-cell compartment itself, although it is these cells that engage in secondary antibody responses capable of mediating immune protection when pre-existing antibodies fail to prevent re-infection. There are few techniques that are capable of detecting rare antigen-specific B cells while also providing information regarding their relative abundance, class/subclass usage and functional affinity. In theory, the ELISPOT/FluoroSpot (collectively ImmunoSpot) assay platform is ideally suited for antigen-specific B-cell assessments since it provides this information at single-cell resolution for individual antibody-secreting cells (ASC). Here, we tested the hypothesis that antigen-coating efficiency could be universally improved across a diverse set of viral antigens if the standard direct (non-specific, low affinity) antigen absorption to the membrane was substituted by high-affinity capture. Specifically, we report an enhancement in assay sensitivity and a reduction in required protein concentrations through the capture of recombinant proteins via their encoded hexahistidine (6XHis) affinity tag. Affinity tag antigen coating enabled detection of SARS-CoV-2 Spike receptor binding domain (RBD)-reactive ASC, and also significantly improved assay performance using additional control antigens. Collectively, establishment of a universal antigen-coating approach streamlines characterization of the memory B-cell compartment after SARS-CoV-2 infection or COVID-19 vaccinations, and facilitates high-throughput immune-monitoring efforts of large donor cohorts in general. Twit Text FOAVip Affinity Tag Coating Enables Reliable Detection of Antigen-Specific B Cells in Immunospot Assays ????Cells http://www.kidney.de/pget.php?&tw=1&pmid=34440612 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34440612 #FOAvip English Wikipedia <ref>{{Cite pmid|34440612}}</ref> Affinity Tag Coating Enables Reliable Detection of Antigen-Specific B Cells in Immunospot Assays #MMPMID34440612 Koppert S; Wolf C; Becza N; Sautto GA; Franke F; Kuerten S; Ross TM; Lehmann PV; Kirchenbaum GA Cells 2021[Jul]; 10 (8): ä PMID34440612show ga Assessment of humoral immunity to SARS-CoV-2 and other infectious agents is typically restricted to detecting antigen-specific antibodies in the serum. Rarely does immune monitoring entail assessment of the memory B-cell compartment itself, although it is these cells that engage in secondary antibody responses capable of mediating immune protection when pre-existing antibodies fail to prevent re-infection. There are few techniques that are capable of detecting rare antigen-specific B cells while also providing information regarding their relative abundance, class/subclass usage and functional affinity. In theory, the ELISPOT/FluoroSpot (collectively ImmunoSpot) assay platform is ideally suited for antigen-specific B-cell assessments since it provides this information at single-cell resolution for individual antibody-secreting cells (ASC). Here, we tested the hypothesis that antigen-coating efficiency could be universally improved across a diverse set of viral antigens if the standard direct (non-specific, low affinity) antigen absorption to the membrane was substituted by high-affinity capture. Specifically, we report an enhancement in assay sensitivity and a reduction in required protein concentrations through the capture of recombinant proteins via their encoded hexahistidine (6XHis) affinity tag. Affinity tag antigen coating enabled detection of SARS-CoV-2 Spike receptor binding domain (RBD)-reactive ASC, and also significantly improved assay performance using additional control antigens. Collectively, establishment of a universal antigen-coating approach streamlines characterization of the memory B-cell compartment after SARS-CoV-2 infection or COVID-19 vaccinations, and facilitates high-throughput immune-monitoring efforts of large donor cohorts in general. |*Immunologic Memory[MESH] |Animals[MESH] |Antigens, Viral/*analysis[MESH] |B-Lymphocytes/*immunology[MESH] |COVID-19[MESH] |Enzyme-Linked Immunospot Assay/*methods[MESH] |Histidine[MESH] |Humans[MESH] |Mice[MESH] |Oligopeptides[MESH] |SARS-CoV-2/*immunology/metabolism[MESH]Affinity Tag Coating Enables Reliable Detection of Antigen-Specific B (epmc).pdf DeepDyve Pubget Overpricing