Multiplex detection of surface molecules on colorectal cancers
(2006) In Proteomics 6(6). p.1791-1802- Abstract
- A technique of fluorescence multiplexing is described for analysis of the plasma membrane proteome of colorectal cancer cells from surgically resected specimens, enabling detection and immunophenotyping when the cancer cells are in the minority. A single-cell suspension was prepared from a colorectal tumour, and the mixed population of cells was captured on a CD antibody microarray. The cancer cells were detected using a fluorescently tagged antibody for carcinoembryonic antigen (CEA-Alexa647) or epithelial cell adhesion marker (EpCAM-Alexa488). Using this multiplexing procedure, dot patterns from colorectal cancers were distinct from those of adjacent normal tissue. Subtraction of the expression levels for each antigen from normal tissue... (More)
- A technique of fluorescence multiplexing is described for analysis of the plasma membrane proteome of colorectal cancer cells from surgically resected specimens, enabling detection and immunophenotyping when the cancer cells are in the minority. A single-cell suspension was prepared from a colorectal tumour, and the mixed population of cells was captured on a CD antibody microarray. The cancer cells were detected using a fluorescently tagged antibody for carcinoembryonic antigen (CEA-Alexa647) or epithelial cell adhesion marker (EpCAM-Alexa488). Using this multiplexing procedure, dot patterns from colorectal cancers were distinct from those of adjacent normal tissue. Subtraction of the expression levels for each antigen from normal tissue from those for the cancer shows differential expression in the cancer of CD66c, CD15s, CD55, CD45, CD71, CD45RO, CD11b and CEA, in descending order. Cells captured on the same microarray were also labelled with fluorescent CD3-phycoerythrin antibody revealing the presence of tumour-infiltrating lymphocytes. The immunophenotypes of T lymphocytes from the tumour samples showed differential expression of HLA-DR, TCR /, CD49d, CD52, CD49e, CD5, CD95, CD28, CD38 and CD71, in descending order. Fluorescence multiplexing of mixed cell populations captured on a single antibody microarray enables expression profiling of multiple sub-populations of cells within a tumour sample. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1059574
- author
- Ellmark, Peter LU ; Belov, Larissa ; Huang, Pauline ; Lee, Soon ; Solomon, Michael ; Morgan, Daniel and Christopherson, Richard
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Proteomics
- volume
- 6
- issue
- 6
- pages
- 1791 - 1802
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:33645472887
- ISSN
- 1615-9861
- DOI
- 10.1002/pmic.200500468
- language
- English
- LU publication?
- no
- id
- 461f7fc4-00f3-4b73-8823-0545f1594487 (old id 1059574)
- date added to LUP
- 2016-04-01 12:35:53
- date last changed
- 2022-03-29 03:03:07
@article{461f7fc4-00f3-4b73-8823-0545f1594487,
abstract = {{A technique of fluorescence multiplexing is described for analysis of the plasma membrane proteome of colorectal cancer cells from surgically resected specimens, enabling detection and immunophenotyping when the cancer cells are in the minority. A single-cell suspension was prepared from a colorectal tumour, and the mixed population of cells was captured on a CD antibody microarray. The cancer cells were detected using a fluorescently tagged antibody for carcinoembryonic antigen (CEA-Alexa647) or epithelial cell adhesion marker (EpCAM-Alexa488). Using this multiplexing procedure, dot patterns from colorectal cancers were distinct from those of adjacent normal tissue. Subtraction of the expression levels for each antigen from normal tissue from those for the cancer shows differential expression in the cancer of CD66c, CD15s, CD55, CD45, CD71, CD45RO, CD11b and CEA, in descending order. Cells captured on the same microarray were also labelled with fluorescent CD3-phycoerythrin antibody revealing the presence of tumour-infiltrating lymphocytes. The immunophenotypes of T lymphocytes from the tumour samples showed differential expression of HLA-DR, TCR /, CD49d, CD52, CD49e, CD5, CD95, CD28, CD38 and CD71, in descending order. Fluorescence multiplexing of mixed cell populations captured on a single antibody microarray enables expression profiling of multiple sub-populations of cells within a tumour sample.}},
author = {{Ellmark, Peter and Belov, Larissa and Huang, Pauline and Lee, Soon and Solomon, Michael and Morgan, Daniel and Christopherson, Richard}},
issn = {{1615-9861}},
language = {{eng}},
number = {{6}},
pages = {{1791--1802}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Proteomics}},
title = {{Multiplex detection of surface molecules on colorectal cancers}},
url = {{http://dx.doi.org/10.1002/pmic.200500468}},
doi = {{10.1002/pmic.200500468}},
volume = {{6}},
year = {{2006}},
}