Methodological comparison of two anti-ZAP-70 antibodies
| Autor: | Heba A. Degheidy, Adrian Wiestner, Wyndham H. Wilson, Gerald E. Marti, M Stetler-Stevenson, Diane C. Arthur, Mohammed Farooqui, Fatima Abbasi, David Venzon |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Male
clone (Java method) Oncology medicine.medical_specialty Histology Scoring system Combined use Gene Expression Biology Article Pathology and Forensic Medicine Antibody Specificity Internal medicine Expression analysis medicine Humans Equivocal result Aged Aged 80 and over ZAP-70 Protein-Tyrosine Kinase Antibodies Monoclonal Cell Biology Middle Aged Flow Cytometry Leukemia Lymphocytic Chronic B-Cell Isotype Improved performance Antigens Surface Immunology biology.protein Female Antibody |
| Zdroj: | Cytometry Part B: Clinical Cytometry. :300-308 |
| ISSN: | 1552-4949 |
| DOI: | 10.1002/cyto.b.20591 |
| Popis: | Background: ZAP-70 expression is a stage independent prognostic marker in CLL. However, interlaboratory variation is large, and there is neither a consensus nor a regulatory approved methodology. Methods: Two anti-ZAP70 clones (1E7.2 and SBZAP) were compared in 45 untreated CLL patients. Nine different methods for ZAP-70 expression analysis were evaluated: M1, isotype control to determine negative; M2, internal residual T-cell to determine positive; M3, normal donor (ND) T-cell to determine positive; M4, internal T-cell/clone ratio; M5, ND residual T-cell/clone ratio; M6, clone/normal remaining B-cell ratio; M7, clone/ND B- cell ratio; M8, CLL-Z score; M9, modified CLL-Z score. A scoring system was designed integrating both 1E7.2 and SBZAP clones to assign ZAP-70 expression. Results: The correlation coefficients for the four selected highest statistically significant methods were as follows (M1 = 0.71, M3 = 0.72, M7 = 0.67, and M9 = 0.64). These four methods were used to generate a combined score. The two reagents showed agreement using the designed scoring system for 37/45 samples (82%), and 8/45 (18%) showed equivocal result with one of the two clones. Seven of the eight equivocal samples were resolved using the scoring system. Conclusions: Four of the nine methods of analysis were compared for each reagent. The use of two independent ZAP-70 reagents increases analytical certitude and the scoring method aids in the resolution of equivocal results. The combined use of two reagents, four methods of analysis, and a scoring method allowed for assignment of ZAP-70 expression in 44/45 samples (98%) tested and improved performance of this important prognostic assay. Published 2011 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text