کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2088277 | 1545717 | 2013 | 7 صفحه PDF | دانلود رایگان |
Autoantibodies to cytokines are important biological effector molecules that can regulate cytokine activities. The aim of the study was to develop a protocol to purify autoantibodies to tumor necrosis factor from human serum, for use as a calibration material to determine the absolute content of autoantibodies to tumor necrosis factor by enzyme-linked immunosorbent assay. The proposed protocol includes a set of affinity chromatography methods, namely, Bio-Gel P6DG sorbent to remove albumin from serum, Protein G Sepharose 4 Fast Flow to obtain a total immunoglobulin G fraction of serum immunoglobulins, and Affi-Gel 15 to obtain specifically antibodies to tumor necrosis factor. The addition of a magnetic separation procedure to the protocol eliminated contaminant tumor necrosis factor from the fraction of autoantibodies to tumor necrosis factor. The protocol generated a pure fraction of autoantibodies to tumor necrosis factor, and enabled us to determine the absolute concentrations of different subclasses of immunoglobulin G autoantibodies to tumor necrosis factor in apparently healthy donors.
► A protocol for affinity purification of IgG autoantibodies to TNF is proposed.
► Magnetic separation is used to remove impurities from TNF.
► The purified fraction of TNF autoantibodies is used for calibration in ELISA.
► We report the absolute contents of different subclasses of IgG autoantibodies to TNF.
Journal: Journal of Immunological Methods - Volume 390, Issues 1–2, 30 April 2013, Pages 92–98