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Improvement of the sensitivity of mono- and bi-dimensional immunoelectrophoretic techniques by transfer onto
S Levasseur1, R Sesboüé, J P Salier
1INSERM Unité 295, U.E.R. de Médecine-Pharmacie, Saint-Etienne du Rouvray, France.
The sensitivity of crossed immunoelectrophoresis (CIE) and rocket immunoelectrophoresis (RIE) does not usually permit the study of diluted protein solutions or the detection of trace components. Antigen-antibody precipitates obtained after immunoelectrophoresis were therefore dissociated and transferred from the agarose gel onto nitrocellulose filter by electrophoresis at pH 11. Immunochemical detection of proteins was then performed with the peroxidase-anti-peroxidase method. Diluted (1/100) solutions gave patterns similar to those obtained with the starting biological fluid. As an example, this technique was applied to human inter-alpha-trypsin inhibitor present at concentrations in the nanogram range. Other proteins, differing in charge and/or molecular weight, were also identified.
The sensitivity of crossed immunoelectrophoresis (CIE) and rocket immunoelectrophoresis (RIE) does not usually permit the study of diluted protein solutions or the detection of trace components. Antigen-antibody precipitates obtained after immunoelectrophoresis were therefore dissociated and transferred from the agarose gel onto nitrocellulose filter by electrophoresis at pH 11. Immunochemical detection of proteins was then performed with the peroxidase-anti-peroxidase method. Diluted (1/100) solutions gave patterns similar to those obtained with the starting biological fluid. As an example, this technique was applied to human inter-alpha-trypsin inhibitor present at concentrations in the nanogram range. Other proteins, differing in charge and/or molecular weight, were also identified.