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目的对特定蛋白检测仪BN-P检测血浆IgG含量的散射比浊法(nephelometry)进行验证,并用于层析法制备IgG的工艺过程监控。方法使用BN-P和IgG相关试剂对检测IgG含量的散射比浊法进行线性和范围、准确度、精密度、专属性、耐用性验证,并进行样品添加试验,将用该方法检测5批IgG成品的结果与凯氏定氮法比较,同时检测层析法制备的3批IgG工艺过程样品中IgG含量。结果用该方法检测蛋白SL标准品中IgG的线性范围为0.107~0.003 34 g/L;高、中、低浓度的SL/H(高水平)蛋白质控品的批内准确度为87.60%~91.05%,批间准确度为88.5%~90.5%;批内精密度CV值为1.85%~3.65%,批间精密度CV值为2.46%~3.16%;检测方法不受血浆白蛋白的干扰,专属性良好;样品经反复冻融5次以内检测,耐受性良好,5次以上耐受性较差;样品添加试验的回收率在90.2%~93.1%之间。该方法检测5批IgG成品中IgG含量,与凯氏定氮法相比差异无统计学意义(P>0.05);该方法检测3批IgG工艺过程中间品重复性较好,步骤回收率一致,工艺稳定。结论特定蛋白检测仪BN-P检测血浆IgG含量的散射比浊法准确可靠,可用于IgG制备工艺的过程监控。
Objective To verify the nephelometry method for the detection of plasma IgG by BN-P in a specific protein detector and to monitor the technological process of the preparation of IgG by chromatography. Methods The linearity, range, accuracy, precision, specificity, durability of the turbidimetric method for the detection of IgG content were tested using BN-P and IgG-related reagents and sample addition tests were performed to detect 5 batches of IgG The final product was compared with the Kjeldahl method, and the IgG content of three batches of IgG process samples prepared by chromatography was also tested. Results The linearity of IgG in protein SL standard was 0.107-0.003 34 g / L. The intra-assay accuracy of high, medium and low concentrations of SL / H protein control was 87.60% -91.05 %, And the inter-assay accuracy was 88.5% -90.5%. The intra-assay CV was 1.85% -3.65%, and the inter-assay CV was 2.46% -3.16%. The detection method was independent of the plasma albumin The samples were tested by repeated freeze-thaw cycles within 5 times. The samples were well tolerated and less than 5 times tolerated. The recoveries of samples were between 90.2% -93.1%. This method was used to detect IgG content in 5 batches of IgG product, which showed no significant difference compared with Kjeldahl method (P> 0.05). This method was better in repeatability of intermediate batches of 3 batches of IgG, stable. Conclusion The specific nephelometry assay of BN-P by plasma-specific turbidimetry can be used to monitor the process of IgG preparation.