Hamilton, A., Alhashimi, F., Myssina, S., Jorgensen, H. G. and Holyoake, T. L. (2009) Optimization of methods for the detection of BCR-ABL activity in Philadelphia-positive cells. Experimental Hematology, 37(3), pp. 395-401. (doi: 10.1016/j.exphem.2008.11.005)
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Publisher's URL: http://dx.doi.org/10.1016/j.exphem.2008.11.005
Abstract
<b>Objective:</b> The recent success in treating chronic myeloid leukemia (CML) with tyrosine kinase inhibitors (TKI), such as imatinib mesylate (IM), has created a demand for reproducible methods to accurately assess inhibition of BCR-ABL activity within CML cells, including rare stein and progenitor cells, either in vitro or in vivo. The purpose of this study was to develop an enzyme-linked immunosorbent (ELISA) method to measure total tyrosine phosphorylation (P-Tyr) in small samples of cells that express BCR-ABL and to compare to more established methods.<p></p> <b>Materials and Methods:</b> The assay was first validated in BCR-ABL wild-type and mutant vs BCR-ABL-negative cell lines. P-Tyr levels were then measured by ELISA in primary CD34(+) CML cells treated with IM.<p></p> <b>Results:</b> In vitro exposure to TKI resulted in decreases in the level of P-Tyr, in both BCR-ABL-positive cell lines and primary CD34(+) CML samples, which were comparable to the reduction in P-Tyr by flow cytometry and phosphorylation of CrkL by either Western blot or flow cytometry.<p></p> <b>Conclusion:</b> We have developed an accurate ELISA method to measure BCR-ABL, activity within Ph+ cells, which is comparable to other in vitro BCR-ABL, assessment techniques in terms of sensitivity and could be adapted for high throughput.<p></p>
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Hamilton, Ms Ashley and Holyoake, Professor Tessa and Jorgensen, Dr Heather |
| Authors: | Hamilton, A., Alhashimi, F., Myssina, S., Jorgensen, H. G., and Holyoake, T. L. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Cancer Sciences |
| Journal Name: | Experimental Hematology |
| Publisher: | Elsevier |
| ISSN: | 0301-472X |
| ISSN (Online): | 1873-2399 |
| Related URLs: |
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