A Simple Method to Overcome the Interference of Hemoglobin in the Detection of Reporter Genes in Vivo

Authors: Reynier P.1; Lesage D.1; Cao A.1; Briane D.1, 3; Coudert R.1, 2; Taillandier E.1

Source: Analytical Biochemistry, Volume 301, Number 2, February 2002 , pp. 217-224(8)

Publisher: Academic Press

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Abstract:

The determination of chemiluminescent intensity of reporter gene expression in vivo is generally disturbed by the presence of hemoglobin. Current methods consist in using perfusion to eliminate blood from investigated tumors or organs. In this work we propose a simple method to overcome this difficulty. The method consists in establishing an absorbance-dependence plot of the ratio R% = Phi/Phi0 between the chemiluminescent intensities measured when hemoglobin is present or absent. For every measurement of the luminescent intensity Phi on sample containing blood, if the absorbance A of the hemoglobin is determined, it allows one to have the intensity ratio R% which in turn gives the corrected intensity Phi0 when the absorption by hemoglobin is eliminated. The method is particularly adapted for comparative measurements of transfection levels in tumors where perfusion cannot be easily performed. ©2002 Elsevier Science (USA).

Keywords: reporter gene; hemoglobin; cationic liposomes; gene therapy

Language: English

Document Type: Research article

Affiliations: 1: UFR de Médecine, Université Paris XIII, 74 rue Marcel Cachin, Bobigny Cedex, F93017, France 2: Faculté des Sciences, Parc de Grandmont, Tours, 37200, France 3: UFR de Médecine, Université Paris XIII, 74 rue Marcel Cachin, Bobigny Cedex, F93017, France

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