Craig W Y, Poulin S E, Nelson C P, Ritchie R F
Foundation for Blood Research, Scarborough, ME 04070-0190.
Clin Chem. 1994 Jun;40(6):882-8.
We describe an ELISA for serum IgG antibodies against malondialdehyde-modified low-density lipoprotein (mLDL). Optimal antigen concentration, serum dilution, and dilution of enzyme-conjugated second antibody were 25 mg/L, 1:250, and 1:5000, respectively, when 5 g/L human serum albumin was used for blocking. When data were expressed as mLDL/LDL (the ratio of IgG binding to mLDL vs LDL), within-run and between-run CVs were 7.0% and 8.9%, respectively. Antibody concentrations expressed as mLDL/LDL or as mLDL-LDL (the difference between IgG binding to mLDL and to LDL) were higher in women with systemic lupus erythematosus (n = 20) than in controls (n = 20) (P < 0.001). With bovine serum albumin or Superblock blocking buffers, only the mLDL-LDL data were significant. Thus, the choice of blocking agent and the method of data expression should be carefully considered when assaying IgG antibodies against mLDL.
我们描述了一种用于检测血清中抗丙二醛修饰低密度脂蛋白（mLDL）IgG抗体的酶联免疫吸附测定（ELISA）法。当使用5g/L人血清白蛋白进行封闭时，最佳抗原浓度、血清稀释度和酶标记二抗的稀释度分别为25mg/L、1:250和1:5000。当数据表示为mLDL/LDL（IgG与mLDL结合量与LDL结合量的比值）时，批内和批间变异系数（CV）分别为7.0%和8.9%。以mLDL/LDL或mLDL-LDL（IgG与mLDL结合量与IgG与LDL结合量的差值）表示的抗体浓度，在系统性红斑狼疮患者（n = 20）中高于对照组（n = 20）（P < 0.001）。使用牛血清白蛋白或Superblock封闭缓冲液时，只有mLDL-LDL数据具有显著性差异。因此，在检测抗mLDL的IgG抗体时，应仔细考虑封闭剂的选择和数据表达方法。
