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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.
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
html.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.


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