A sensitive solid phase microradioimmunoassay for anti-double stranded DNA antibodies

Abstract

A sensitive solid phase microradioimmunoassay has been developed for measurement of antidouble stranded DNA (dsDNA) antibodies. In this procedure, advantage has been taken of the capacity of poly-L-lysine (PLL) to facilitate the binding of pure dsDNA to plastic surfaces. In the absence of PLL, binding did not occur. Diluted sera were incubated in PLL-treated dsDNA-coated microtitration trays and anti-dsDNA Ig measured using affinity purified 125I-anti-Ig of high specific activity. The synthetic DNA, poly dA-dT, was used as a model for dsDNA. In initial experiments, specific anti-DNA binding could not be demonstrated because of high background binding of patient Ig to PLL-treated surfaces. This was reduced by diluting test sera and anti-Ig in buffer containing 2% BGG and 1% BSA. Specificity of the assay for DNA was demonstrated by absorbing the anti-DNA activity on DNA-coated plastic. The binding of systemic lupus erythematosus (SLE) patient serum Ig to poly dA-dT coated trays did not diminish after digestion with nuclease S1, suggesting that the synthetic polymer is an appropriate model for dsDNA. Patient and normal sera were screened for anti-dsDNA activity using poly dA-dT as antigen. None of the 38 normal sera, 23 of 35 active SLE sera, 1 of 25 treated SLE, 4 of 35 rheumatoid arthritis, 3 of 35 scleroderma, and 1 of 13 polymyositis sera demonstrated positive anti-dsDNA activity. The anti-dsDNA values obtained in the radioimmunoassay correlated significantly with those obtained in the Crithidia luciliae assay.