Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides-2

Or that donor)×100. This allowed SD to be compared on the same scale. Note that the regression line for ELISPOT (dotted) is consistently higher than the regression line for CFC (dashed) or tetramer (solid). Due to complexity, data points for different donors and different antigens are not distinguished in this Figure; please refer to Figure 2 for relative responses of individual donors and antigens.<p><b>Copyright information:</b></p><p>Taken from "Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides"</p><p>http://www.biomedcentral.com/1471-2172/9/9</p><p>BMC Immunology 2008;9():9-9.</p><p>Published online 17 Mar 2008</p><p>PMCID:PMC2275721.</p><p></p

Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides-3

Onder. The same non-responding donor was used for all assays. Unstimulated background was subtracted for each dilution point in CFC and ELISPOT assays. This background was uniformly low (< 0.08% for CFC and < 15 SFC per 2.5 × 10PBMC for ELISPOT) despite the used of allogeneic PBMC for the dilution. Note that the pp65peptide response of this donor (#43) was much lower in ELISPOT compared to the other two assays.<p><b>Copyright information:</b></p><p>Taken from "Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides"</p><p>http://www.biomedcentral.com/1471-2172/9/9</p><p>BMC Immunology 2008;9():9-9.</p><p>Published online 17 Mar 2008</p><p>PMCID:PMC2275721.</p><p></p

Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides-0

Nd ELISPOT assays. Circles represent donor 41; triangles, donor 68; and squares, donor 43. Open symbols represent CMV pp65responses; closed symbols represent CMV pp65 peptide mix responses (CFC and ELISPOT assays only). Note that certain responses were very similar, so some symbols overlap. Error bars represent the SD of 10 times that the six replicates were repeated. The gray zones indicate the area within which a laboratory doing validation could expect their data to lie.<p><b>Copyright information:</b></p><p>Taken from "Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides"</p><p>http://www.biomedcentral.com/1471-2172/9/9</p><p>BMC Immunology 2008;9():9-9.</p><p>Published online 17 Mar 2008</p><p>PMCID:PMC2275721.</p><p></p

Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides-1

Ys on different days (inter-assay precision), or three operators on the same day (inter-operator precision). Circles represent donor 41; triangles, donor 68; and squares, donor 43. Open symbols represent CMV pp65responses; closed symbols represent CMV pp65 peptide mix responses (CFC and ELISPOT assays only). Error bars in the intra-assay graphs represent the SD of 10 times that the six replicates were repeated. Lines represent linear regression of the combined data (both antigens, where used), with 95% confidence intervals of the regression shown with dotted lines.<p><b>Copyright information:</b></p><p>Taken from "Precision and linearity targets for validation of an IFNγ ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides"</p><p>http://www.biomedcentral.com/1471-2172/9/9</p><p>BMC Immunology 2008;9():9-9.</p><p>Published online 17 Mar 2008</p><p>PMCID:PMC2275721.</p><p></p