Rauch J, Massicotte H, Wild J, Tannenbaum H
J Rheumatol. 1985 Jun;12(3):482-6.
Serum from patients with systemic lupus erythematosus (SLE) and hybridoma culture fluids derived from the fusion of SLE lymphocytes contain antibodies to native DNA (nDNA) and denatured DNA (dDNA). A rapid, efficient solid phase radioimmunoassay (RIA) was developed to screen for minute quantities of these autoantibodies. The RIA, which utilized polystyrene tubes, required the addition of 0.1% bovine serum albumin and 0.01% Tween 20 detergent to decrease nonspecific immunoglobulin binding. Pretreatment of the polystyrene tubes with poly-L-lysine (PLL) prior to coating with DNA increased the binding of radiolabeled nDNA from 15 to 46% and of dDNA from 17 to 63%. This PLL precoating step resulted in a 3-fold increase in the specificity of the assay for nDNA but was not advantageous for dDNA. The method described is sensitive, specific, and can be applied to the screening of microgram quantities of anti-DNA autoantibodies in serum and hybridoma culture fluids.
系统性红斑狼疮（SLE）患者的血清以及源自SLE淋巴细胞融合的杂交瘤培养液中含有抗天然DNA（nDNA）和变性DNA（dDNA）的抗体。开发了一种快速、高效的固相放射免疫测定法（RIA）来筛查微量的这些自身抗体。该RIA使用聚苯乙烯管，需要添加0.1%的牛血清白蛋白和0.01%的吐温20去污剂以减少非特异性免疫球蛋白结合。在用DNA包被之前，用聚-L-赖氨酸（PLL）预处理聚苯乙烯管，使放射性标记的nDNA结合从15%增加到46%，dDNA结合从17%增加到63%。这一PLL预包被步骤使该测定法对nDNA的特异性提高了3倍，但对dDNA并无优势。所描述的方法灵敏、特异，可用于筛查血清和杂交瘤培养液中微克量的抗DNA自身抗体。
