A High-Sensitivity Method for Detection and Measurement of HMGB1 Protein Concentration by High-Affinity Binding to DNA Hemicatenanes
Figure 4
HMGB1 assay in serums from patients.
(A) without PCA treatment, or (B) after sample purification by PCA treatment. On each gel the five samples on the left, prepared with known amounts of pure HMGB1, were used to calibrate the method. The 15 serums tested are labeled a-o, serum a originating from a healthy individual, serums b-o from randomly selected patients in intensive care unit (d,g,h,i,k,l,n: sepsis; b,c,e,f,j,m,o: septic shock). Note the lower sensitivity, higher background, and non-specific bands (arrowheads) obtained with crude serums, as compared with the strong increase in sensitivity and marked background decrease after treatment of serums with PCA. (C) Comparison of band-shift assay with ELISA. Concentrations measured by ELISA are plotted against band-shift assay data obtained after PCA treatment of serums, on a double logarithmic scale. Each circled letter on the diagram represents the corresponding serum as assayed in (B).