Determination of Nerve Growth Factor in Rat Spinal Cord by Capillary Electrophoresis-Based Immunoassay with a Laser Induced Fluorescence Detector

Abstract

In order to study the relationship between spinal cord injury and the change of nerve growth factor (NGF), an analytical method for NGF by capillary electrophoresis-based immunoassay (CEIA) with laser-induced fluorescence (LIF) was developed. Having been dissolved in phosphate buffer solution and concentrated with vacuum freeze-drying, NGF in spinal cord of rat was allowed to react with NGF monoclonal antibody labeled with fluorescence isothiocyanate (FITC). Then the immuno-complex, FITC-labeled anti-NGF and FITC were separated and determined by LIF-CEIA using Kiton red as the internal standard. The linear range of the method was 2–30 ng mL−1 and the limit of detection was 0.35 ng mL−1. The relative standard derivations for relative migration time and relative fluorescence intensity ratio were 7.98% and 6.52%, respectively. The contents of NGF from spinal cord of rat were determined by both the proposed method and Western blotting. The results with the two methods agreed well. The spiked recoveries of the samples were 88.5–116.3%. The proposed method was rapid, precise and inexpensive.