Immunodetection and molecular forms of plasma vascular endothelial growth factor-C.

2.50
Hdl Handle:
http://hdl.handle.net/10541/79114
Title:
Immunodetection and molecular forms of plasma vascular endothelial growth factor-C.
Authors:
Duff, Sarah E; Li, Chenggang; Renehan, Andrew G; O'Dwyer, Sarah T; Kumar, Shant
Abstract:
Vascular endothelial growth factor (VEGF)-C is a member of the VEGF family. VEGF-C is involved in developmental lymphangiogenesis and may be important in pathological lymphangiogenesis, lymphatic invasion and metastasis in carcinoma. We describe the development of an indirect enzyme-linked immunosorbent (ELISA) assay for the quantification of VEGF-C in plasma. Capture of VEGF-C was achieved using goat anti-human VEGF-C antibody, followed by detection with rabbit anti-human VEGF-C antibody. The sensitivity of the assay was amplified using the biotin-avidin and enhanced chemiluminescence (ECL) systems. The assay was highly sensitive and reproducible with a detection range of 0.4-100 U/ml and the intra- and inter-assay variations were less than 8%. Substitutional tests demonstrated that the assay was specific for VEGF-C with no cross-reaction with VEGF-A or VEGF-D. Practical application of the assay was evaluated in 41 colorectal cancer patients and 31 controls. Median plasma levels of VEGF-C were 35.0 U/ml (range: 17.4-75.9 U/ml) in colorectal cancer patients in contrast to 11.5 U/ml (range: 5.4-21.5 U/ml) in controls (p<0.001). Moreover, VEGF-C levels tended to be elevated in patients with advanced disease compared to early disease, but this was not statistically significant owing to a relatively small number of patients in each group. Immunoprecipitation and immunoblotting confirmed detection of VEGF-C in plasma and revealed that two forms of VEGF-C were present in the plasma corresponding to approximately 40 and approximately 80 kDa. The measurement of plasma VEGF-C offers opportunities to explore clinical applications in the management of malignancy, in particular in the prediction of lymphatic spread and in other lymphangiogenesis-related diseases.
Affiliation:
Laboratory Medicine Academic Group, Department of Experimental Pathology, Stopford Building, University of Manchester, Manchester M13 9PT, UK. sarah.duff@man.ac.uk
Citation:
Immunodetection and molecular forms of plasma vascular endothelial growth factor-C. 2003, 22 (2):339-43 Int. J. Oncol.
Journal:
International Journal of Oncology
Issue Date:
Feb-2003
URI:
http://hdl.handle.net/10541/79114
PubMed ID:
12527932
Type:
Article
Language:
en
ISSN:
1019-6439
Appears in Collections:
All Christie Publications

Full metadata record

DC FieldValue Language
dc.contributor.authorDuff, Sarah E-
dc.contributor.authorLi, Chenggang-
dc.contributor.authorRenehan, Andrew G-
dc.contributor.authorO'Dwyer, Sarah T-
dc.contributor.authorKumar, Shant-
dc.date.accessioned2009-08-28T11:37:48Z-
dc.date.available2009-08-28T11:37:48Z-
dc.date.issued2003-02-
dc.identifier.citationImmunodetection and molecular forms of plasma vascular endothelial growth factor-C. 2003, 22 (2):339-43 Int. J. Oncol.en
dc.identifier.issn1019-6439-
dc.identifier.pmid12527932-
dc.identifier.urihttp://hdl.handle.net/10541/79114-
dc.description.abstractVascular endothelial growth factor (VEGF)-C is a member of the VEGF family. VEGF-C is involved in developmental lymphangiogenesis and may be important in pathological lymphangiogenesis, lymphatic invasion and metastasis in carcinoma. We describe the development of an indirect enzyme-linked immunosorbent (ELISA) assay for the quantification of VEGF-C in plasma. Capture of VEGF-C was achieved using goat anti-human VEGF-C antibody, followed by detection with rabbit anti-human VEGF-C antibody. The sensitivity of the assay was amplified using the biotin-avidin and enhanced chemiluminescence (ECL) systems. The assay was highly sensitive and reproducible with a detection range of 0.4-100 U/ml and the intra- and inter-assay variations were less than 8%. Substitutional tests demonstrated that the assay was specific for VEGF-C with no cross-reaction with VEGF-A or VEGF-D. Practical application of the assay was evaluated in 41 colorectal cancer patients and 31 controls. Median plasma levels of VEGF-C were 35.0 U/ml (range: 17.4-75.9 U/ml) in colorectal cancer patients in contrast to 11.5 U/ml (range: 5.4-21.5 U/ml) in controls (p<0.001). Moreover, VEGF-C levels tended to be elevated in patients with advanced disease compared to early disease, but this was not statistically significant owing to a relatively small number of patients in each group. Immunoprecipitation and immunoblotting confirmed detection of VEGF-C in plasma and revealed that two forms of VEGF-C were present in the plasma corresponding to approximately 40 and approximately 80 kDa. The measurement of plasma VEGF-C offers opportunities to explore clinical applications in the management of malignancy, in particular in the prediction of lymphatic spread and in other lymphangiogenesis-related diseases.en
dc.language.isoenen
dc.subjectColorectal Canceren
dc.subjectCancer Proteinsen
dc.subjectBiological Tumour Markersen
dc.subject.meshAdenocarcinoma-
dc.subject.meshAnimals-
dc.subject.meshAvidin-
dc.subject.meshBiotin-
dc.subject.meshChemiluminescent Measurements-
dc.subject.meshColorectal Neoplasms-
dc.subject.meshCross Reactions-
dc.subject.meshEndothelial Growth Factors-
dc.subject.meshEnzyme-Linked Immunosorbent Assay-
dc.subject.meshFluorescent Antibody Technique, Indirect-
dc.subject.meshGoats-
dc.subject.meshHumans-
dc.subject.meshNeoplasm Proteins-
dc.subject.meshProtein Isoforms-
dc.subject.meshRabbits-
dc.subject.meshRecombinant Proteins-
dc.subject.meshReproducibility of Results-
dc.subject.meshSensitivity and Specificity-
dc.subject.meshSpecies Specificity-
dc.subject.meshTumor Markers, Biological-
dc.subject.meshVascular Endothelial Growth Factor C-
dc.titleImmunodetection and molecular forms of plasma vascular endothelial growth factor-C.en
dc.typeArticleen
dc.contributor.departmentLaboratory Medicine Academic Group, Department of Experimental Pathology, Stopford Building, University of Manchester, Manchester M13 9PT, UK. sarah.duff@man.ac.uken
dc.identifier.journalInternational Journal of Oncologyen

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