Preparation of Monoclonal Antibody Against Chicken Apolipoprotein B and Development of Enzyme Liked Immunosolvent Assay (ELISA) Method with the Antibody Aiming at the Optimization of Lipid Metabolism in Chickens

Abstract

Apolipoprotein B (apoB) containing lipoproteins are major lipoproteins in blood and the elevated concentration in plasma may lead to increased transport of lipoproteins into adipose tissues resulting in the excessive fat deposition. Aims of the present study were to prepare monoclonal antibodies against chicken apoB and to development enzyme linked mmunosolvent assay (ELISA) methods with the antibody which would be provided for studies on lipoprotein metabolism in chickens. A monoclonal antibody, CAB4, was prepared using a purified chicken apoB and specific reactivity of the antibody was verified by Western blot analysis and immuno-precipitation of apoB in plasma and primary hepatocyte cultures in chickens. ELISA method with monoclonal anti-chicken apoB antibody developed in this study was applicable to measure apoB concentration in plasma and cell cultures within the range of 0 to 500ng/ml with the intra- and inter-assay variation of less than 10.6%. Using the present ELISA method, plasma apoB concentrations in broiler chickens starved for 8h were shown to be 0.44mg/ml against 1.02mg/ml in fed chicken counterparts. The monoclonal antibodies against chicken apoB and ELISA method developed in the present study could be potentially used for studies on the lipoprotein metabolism focusing on the optimization of lipid metabolism in chickens.

Keywords

• apolipoprotein b
• monoclonal antibody
• enzyme linked immunosorbent assay
• fat deposition
• broiler chickens