Capurro M, Wanless IR, Sherman M, et al
Capurro M, Wanless IR, Sherman M, et al. detectable concentration was 0.05?ng/ml; the intraassay coefficient of variation (CV) and interassay Mouse monoclonal to CD62L.4AE56 reacts with L-selectin, an 80 kDaleukocyte-endothelial cell adhesion molecule 1 (LECAM-1).CD62L is expressed on most peripheral blood B cells, T cells,some NK cells, monocytes and granulocytes. CD62L mediates lymphocyte homing to high endothelial venules of peripheral lymphoid tissue and leukocyte rollingon activated endothelium at inflammatory sites CV were both less than 10%, with good stability and reproducibility. GPC3 has a high sensitivity (54.2%) and specificity (99.4%) in diagnosing HCC. The level of GPC3 in HCC was robust higher than that in healthy or other liver diseases’ sera (108.67 230.04?ng/ml vs. 3.99 7.68?ng/ml). The diagnostic sensitivity of GPC3 single or combined with CK19 and AFP for HCC was evaluated, and the rates were 54.2 and 90.6%, respectively. Conclusions An applicable chemiluminescent immunoassay with stable performance against GPC3 in diagnosing HCC has been established and the combination SU14813 of GPC3 with CK19 and AFP could improve the diagnostic sensitivity for HCC. axis) and the luminescence value as ordinate (axis). A total of 22 points (concentration range: 0C7,000?ng/ml) was selected to evaluate the reagent’s hook effect. Analysis of the accuracy The analytical accuracy was evaluated by added recovery and dilution recovery. The GPC3 antigen was added to normal human serum with the theoretical concentrations 2, 5, 50, 100, 200, 300, and 500 ng/mL. SU14813 The dilution samples were prepared by 4,000, 2,000, and 1,000?ng/ml and were diluted twofold, fivefold, and tenfold, respectively. Then, the ratio between the measured and theoretical values (the ratio should be between 90.00 and 110.0%) was calculated. Analysis of the minimum detectable concentration Zero reference standard was measured 20 times, the mean and standard deviation (SD) were calculated and substituted with SD into the standard curve equation to obtain the minimum detectable concentration. Specificity test The cross\reactivity of SU14813 this method with several tumor markers associated with HCC, including AFP (1,000?ng/ml, from the AFP time\resolved immunofluorescence assay kit, cat number: DR\CM\A001), CEA (carcinoembryonic antigen; 1,000?ng/ml, from Meridian Life Science, Inc., catalog number: A32321H), and CK19 (800 U/ml, from the CK19 time\resolved immunofluorescence assay kit, cat number: DR\CM\A012), added these markers to normal human serum, respectively; then the luminescence values were measured. Interfering test The interference immunity of the CLIA reagents developed in this experiment was added to the common serum constituents known as causing interference in immunoassays (hemolytic, high cholesterol, high bilirubin). The human hemoglobin (1,000, 500?ng/ml), triglycerides (1,000, 500?ng/ml), and bilirubin (50, 25?ng/ml) were added to the GPC3 antigen samples, respectively. The GPC3 antigen was diluted to 50, 100, and 500?ng/ml for the test. The above solutions were tested and the recovery rate was calculated. Test of stability GPC3 reagent was placed at 37C incubator for 7 days or 4C for 6 months. Then, the physical appearance was observed and the performance was detected during various stages. Detection of clinical specimens One hundred ninety\two patients with HCC, 57 with hepatitis, 54 with gastric cancer, 92 with colorectal cancer, 31 with esophageal cancer, 44 with liver cirrhosis, and 48 cases of healthy human serum samples were evaluated using the GPC3\CLIA reagent. Determination of serum AFP and CK19 Expression of AFP in clinical serum samples was tested by AFP time\resolved detection kit. The CK19 was detected by CK19 time\resolved detection kit, operated according to the manufacturer’s instructions. Statistical methods Statistical analysis of the results was performed using SPSS 13.0. The 0.05. RESULTS Results of Paired Antibody Screening Thirteen GPC3 mcAbs were cycling paired, it was found that the highest fluorescence efficiency was produced using the 8G6 mcAb as the coated antibody and 7D11 mcAb as the detection antibody (Fig.?1). Six series GPC3 antigen concentrations, 0, 31.25, 62.5, 125, 250, and 500 ng/mL, were labeled as standard. The antigen concentration as the abscissa (axis) and luminescence value as vertical axis (axis) showed the matching antibodies can obtain a good linear.