| Zugriffsnummer | 33706 |
| Dokumenttyp | Zeitschriftenartikel |
| Peer Review | unbekannt |
| Sprache | Englisch |
| Titel | Quantification of cells with specific phenotypes I: Determination of CD4+ cell count per microliter in reconstituted lyophilized human PBMC prelabeled with anti-CD4 FITC antibody |
| Autor(in); Institution |
Wang, L.; Biosystems and Biomaterials Division,National Institute of Standards and Technology (NIST), Gaithersburg, USA
Stebbings, R.; Biotherapeutics Group, National Institute for Biological Standards and Control (NIBSC), Potters Bar, UNITED KINGDOM
Gaigalas, A.; Biosystems and Biomaterials Division,National Institute of Standards and Technology (NIST), Gaithersburg, USA
Sutherland, J.; Biotherapeutics Group, National Institute for Biological Standards and Control (NIBSC), Potters Bar, UNITED KINGDOM
Kammel, Martin; 8.3, Biomedizinische Optik, PTB-Berlin
John, Manuela; 8.3, Biomedizinische Optik, PTB-Berlin
Roemer, B.; Abbott GmbH & Co. KG, Wiesbaden, GERMANY
Kuhne, M.; Department of Analytical Chemistry, BAM Federal Institute for Materials Research and Testing, Berlin, GERMANY
Schneider, R.; Department of Analytical Chemistry, BAM Federal Institute for Materials Research and Testing, Berlin, GERMANY
Braun, M.; Beckman Coulter GmbH, Krefeld, GERMANY
Engel, A.; Becton Dickinson GmbH, Heidelberg, GERMANY
Dikshit, D.; Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute (CDRI), Lucknow, INDIA
Abbasi, F.; Center for Devices and Radiologic Health, Food and drug Administration (CDRH/FDA), Bethesda, USA
Marti, G.; Center for Devices and Radiologic Health, Food and drug Administration (CDRH/FDA), Bethesda, USA
Sassi, M.; Amount of Substance, Istituto Nazionale di Ricerca Metrologica (INRIM), Torino, ITALY
Revel, L.; Amount of Substance, Istituto Nazionale di Ricerca Metrologica (INRIM), Torino, ITALY
Kim, S.; Bioanalysis, Korea Research Institute of Standards and Science (KRISS), Daejeon, KOREA
Baradez, M.; Science and Innovation, LGC Limited, Teddington, UNITED KINGDOM
Lekishvili, T.; Science and Innovation, LGC Limited, Teddington, UNITED KINGDOM
Marshall, D.; Science and Innovation, LGC Limited, Teddington, UNITED KINGDOM
Whitby, L.; UK National External Quality Assessment Service (UK NEQAS) for Leucocyte Immunophenotyping, Sheffield, UNITED KINGDOM
Jing, W.; Division of Medical and Biological Measurement, National Institute of Metrology (NIM), Beijing, CHINA
Ost, V.; Partec GmbH, Muenster, GERMANY
Vonsky, M.; Department of State Standards in the Field of Physical - Chemical Measurements, D.I. Mendeleev Institute for Metrology (VNIIM), St-Petersburg, RUSSIA
Neukammer, Jörg; 8.3, Biomedizinische Optik, PTB-Berlin
|
| Quelle/Jahr | Cytometry A: 87 (2015), 3, 244 - 253 |
| ISSN | 0196-4763 (PRINT) ; 1552-4922 (ONLINE) |
| DOI | |
| Verlag | Hoboken, NJ: Wiley Blackwell |
| Freie Schlagworte | CD41 cell counting, relative concentration measurement ; lyophilized cells, flow cytometry ; standard measurement procedure ; measurement of uncertainty ; acquired immunodeficiency syndrome ; reference material ; human immunodeficiency virus-1 |
| Zusammenfassung | A surface-labeled lyophilized lymphocyte (sLL) preparation has been developed using human peripheral blood mononuclear cells prelabeled with a fluorescein isothiocyanate conjugated anti-CD4 monoclonal antibody. The sLL preparation is intended to be used as a reference material for CD41 cell counting including the development of higher order reference measurement procedures and has been evaluated in the pilot study CCQM-P102. This study was conducted across 16 laboratories from eight countries to assess the ability of participants to quantify the CD41 cell count of this reference material and to document cross-laboratory variability plus associated measurement uncertainties. Twelve different flow cytometer platforms were evaluated using a standard protocol that included calibration beads used to obtain quantitative measurements of CD41 T cell counts. There was good overall cross-platform and counting method agreement with a grand mean of the laboratory calculated means of (301.7 +- 4.9) µL-1 CD41 cells. Excluding outliers, greater than 90% of participant data agreed within +-15%. A major contribution to variation of sLL CD41 cell counts was tube to tube variation of the calibration beads, amounting to an uncertainty of 3.6%. Variation due to preparative steps equated to an uncertainty of 2.6%. There was no reduction in variability when data files were centrally reanalyzed. Remaining variation was attributed to instrument specific ifferences. CD41 cell counts obtained in CCQM-P102 are in excellent agreement and show the robustness of both the measurements and the data analysis and hence the suitability of sLL as a reference material for interlaboratory comparisons and external quality assessment. |
Zitierung
Wang, L., Stebbings, R., Gaigalas, A., Sutherland, J., Kammel, M., John, M., Roemer, B., Kuhne, M., Schneider, R., Braun, M., Engel, A., Dikshit, D., Abbasi, F., Marti, G., Sassi, M., Revel, L., Kim, S., Baradez, M., Lekishvili, T., Marshall, D., Whitby, L., Jing, W., Ost, V., Vonsky, M., & Neukammer, J. (2015). Quantification of cells with specific phenotypes I: Determination of CD4+ cell count per microliter in reconstituted lyophilized human PBMC prelabeled with anti-CD4 FITC antibody. Cytometry A, 87(3), 244–253. https://doi.org/10.1002/cyto.a.22634