TY - JOUR

T1 - Development of an ELISA for the quantification of fibrin in canine tumours

AU - Raut, Sanjeev

AU - McEvoy, Fintan

AU - Gaffney, Patrick J.

N1 - Funding Information: The authors thank Joanna McNab for her assistance with the statistical evaluation of the data. This work was supported by a grant from the D. Collen Research Foundation, University of Leuven, Leuven, Belgium.

PY - 1999/10/1

Y1 - 1999/10/1

N2 - Fibrin is found in most solid tumours, and there is speculation regarding its role in tumour invasion and metastasis. An assay to quantitate fibrin levels in tissues would be a useful preliminary tool in assessing the above. Such an assay to quantitatively detect levels of fibrin in various types of canine tumour was developed. This procedure involved an ELISA using a monoclonal antibody (MAb 1H10) for canine fibrin as a capture antibody and a polyclonal antibody to human fibrinogen conjugated to horseradish peroxidase as the detection antibody. The ELISA is calibrated against known concentrations of freeze-fractured fibrin derived from clotted dog plasma. The assay takes 3.5 hours, and concentrations as low as 0.1 μg fibrin per milliliter of solubilised tumour can be readily detected. ELISA dilution curves for fibrin from various types of canine tumour were found to be parallel to the standard canine fibrin calibration curve. The intra-assay and interassay variabilities of the assay gave coefficients of variation in the range of 2.4-4.5% and 7.2-7.8%, respectively, for the calibrator standard, in a concentration range of 0.1-10 μg/ml. Using this assay, we reported the levels of fibrin in three different types of malignant canine tissue. Copyright (C) 1999 Elsevier Science Ltd.

AB - Fibrin is found in most solid tumours, and there is speculation regarding its role in tumour invasion and metastasis. An assay to quantitate fibrin levels in tissues would be a useful preliminary tool in assessing the above. Such an assay to quantitatively detect levels of fibrin in various types of canine tumour was developed. This procedure involved an ELISA using a monoclonal antibody (MAb 1H10) for canine fibrin as a capture antibody and a polyclonal antibody to human fibrinogen conjugated to horseradish peroxidase as the detection antibody. The ELISA is calibrated against known concentrations of freeze-fractured fibrin derived from clotted dog plasma. The assay takes 3.5 hours, and concentrations as low as 0.1 μg fibrin per milliliter of solubilised tumour can be readily detected. ELISA dilution curves for fibrin from various types of canine tumour were found to be parallel to the standard canine fibrin calibration curve. The intra-assay and interassay variabilities of the assay gave coefficients of variation in the range of 2.4-4.5% and 7.2-7.8%, respectively, for the calibrator standard, in a concentration range of 0.1-10 μg/ml. Using this assay, we reported the levels of fibrin in three different types of malignant canine tissue. Copyright (C) 1999 Elsevier Science Ltd.

KW - Assay

KW - ELISA

KW - Fibrin

KW - Monoclonal antibody

KW - Tumour

UR - http://www.scopus.com/inward/record.url?scp=0032729569&partnerID=8YFLogxK

U2 - 10.1016/S0049-3848(99)00054-7

DO - 10.1016/S0049-3848(99)00054-7

M3 - Journal article

C2 - 10554080

AN - SCOPUS:0032729569

VL - 96

SP - 11

EP - 17

JO - Thrombosis Research

JF - Thrombosis Research

SN - 0049-3848

IS - 1

ER -