A Novel Method for Real-Time, Continuous, Fluorescence-Based Analysis of Anti-DNA Abzyme Activity in Systemic Lupus

<table>Continuous measurement of oligo 18-mer hydrolysis by lupus patient (LP) versus normal healthy donor (ND) by fluorescence assay. *Comparison of lupus patient-derived anti-poly-(dT) ssDNA antibody (LP4) and normal healthy donor-derived anti-poly-(dT) ssDNA antibody (NDX) via continuous fluorescence-based assay on Fluoroskan II (Finstruments) demonstrates that 5′-6-FAM-ATATAGCGC<sub>5</sub>T<sub>5</sub>-DQ1-3′ 18-mer hydrolysis probe is cleaved by lupus patient-derived antibody but not by normal donor-derived antibody. Oligo blank contained all reaction components except for antibody (25 mM Tris-HCl, 50 mM NaCl, 10 mM MgCl<sub>2</sub>, pH 7.5, and 1 <i>μ</i>g/mL 5′-6-FAM-ATATAGCGC<sub>5</sub>T<sub>5</sub>-DQ1-3′ 18-mer hydrolysis probe). ANOVA indicates all three fluorescence patterns of LP4, NDX, and the oligo control are significantly different from each other at <svg height="11.375" id="M4" style="vertical-align:-0.1638pt" version="1.1" viewbox="0 0 75.025002 11.375" width="75.025002" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
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Autoimmune Diseases

fig6

Figure 6

Figure 6: A Novel Method for Real-Time, Continuous, Fluorescence-Based Analysis of Anti-DNA Abzyme Activity in Systemic Lupus 