目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO...目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO稀释液、标准对照材料浸提液、56批真实世界的医疗器械产品,从简单到复杂分别比较RTCA法与MTT法用于细胞毒性检测结果的一致性,并且汇总多次试验的阴性对照和阳性对照结果,比较两种方法的可重复性。结果RTCA法检测不同浓度的DMSO稀释液没有剂量效应关系,检测标准对照材料浸提液与MTT法结果完全一致并高度相关(R=0.9898),检测56批医疗器械产品与MTT法结果一致率为80.4%,仅中度相关(R=0.6183)。RTCA法的可重复性不比MTT法好。结论RTCA法能有效检测医疗器械细胞毒性,但是部分产品可能因溶出物干扰检测结果而不适用。RTCA法作为一种新型的医疗器械细胞毒性检测方法的适用性还需要进一步研究。展开更多
We label-free and real-time detected three interaction processes of antigen-antibodies,Human Immunoglobulin G(IgG),Rabbit IgG,and Mouse IgG as the targets,and Goat Anti-human IgG,Goat Anti-rabbit IgG,and Goat Anti-mou...We label-free and real-time detected three interaction processes of antigen-antibodies,Human Immunoglobulin G(IgG),Rabbit IgG,and Mouse IgG as the targets,and Goat Anti-human IgG,Goat Anti-rabbit IgG,and Goat Anti-mouse IgG as the probe,by the Oblique-incidence Reflectivity Difference(OIRD) method.The interaction dynamic curves of the OIRD signal,corresponding to the interaction processes of antigen-antibodies,are generated.The reaction times from beginning to equilibrium state are about 1800,900,and 1200 s for Human IgG,Rabbit IgG,and Mouse IgG,respectively.The experimental results demonstrate that the OIRD method not only can distinguish biomolecular interactions,but also can be used in real-time detection of interactions and dynamic processes of biomolecules.展开更多
Selected Mouse IgG of 1 mg/mL as target was fabricated on microarray for 500 sample dots.Label-free and real-time reaction dynamic processes were detected between the microarrays with Goat Anti-mouse IgG of 0.02 mg/mL...Selected Mouse IgG of 1 mg/mL as target was fabricated on microarray for 500 sample dots.Label-free and real-time reaction dynamic processes were detected between the microarrays with Goat Anti-mouse IgG of 0.02 mg/mL using the obliqueincidence reflectivity difference(OIRD)method.We obtained the reaction results and the reaction dynamic curves of 500 protein dots.In addition,we also used label-free detection of protein microarrays of 10080 sample dots,including BSA and different concentrations of Mouse IgG and Rabbit IgG,by OIRD.The obtained reaction results between the protein microarray with 1 mg/mL Goat Anti-mouse IgG and 1 mg/mL Goat Anti-rabbit IgG are reported herein.Experimental results show that OIRD can be not only label-free high-throughput detection method for biological microarrays but also label-free real-time detection in the interaction processes of biomolecules.展开更多
文摘目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO稀释液、标准对照材料浸提液、56批真实世界的医疗器械产品,从简单到复杂分别比较RTCA法与MTT法用于细胞毒性检测结果的一致性,并且汇总多次试验的阴性对照和阳性对照结果,比较两种方法的可重复性。结果RTCA法检测不同浓度的DMSO稀释液没有剂量效应关系,检测标准对照材料浸提液与MTT法结果完全一致并高度相关(R=0.9898),检测56批医疗器械产品与MTT法结果一致率为80.4%,仅中度相关(R=0.6183)。RTCA法的可重复性不比MTT法好。结论RTCA法能有效检测医疗器械细胞毒性,但是部分产品可能因溶出物干扰检测结果而不适用。RTCA法作为一种新型的医疗器械细胞毒性检测方法的适用性还需要进一步研究。
基金supported by the National Basic Research Program of China (Grant No. 2007CB935701)
文摘We label-free and real-time detected three interaction processes of antigen-antibodies,Human Immunoglobulin G(IgG),Rabbit IgG,and Mouse IgG as the targets,and Goat Anti-human IgG,Goat Anti-rabbit IgG,and Goat Anti-mouse IgG as the probe,by the Oblique-incidence Reflectivity Difference(OIRD) method.The interaction dynamic curves of the OIRD signal,corresponding to the interaction processes of antigen-antibodies,are generated.The reaction times from beginning to equilibrium state are about 1800,900,and 1200 s for Human IgG,Rabbit IgG,and Mouse IgG,respectively.The experimental results demonstrate that the OIRD method not only can distinguish biomolecular interactions,but also can be used in real-time detection of interactions and dynamic processes of biomolecules.
基金supported by the National Basic Research Program of China(Grant No.2007CB35710)
文摘Selected Mouse IgG of 1 mg/mL as target was fabricated on microarray for 500 sample dots.Label-free and real-time reaction dynamic processes were detected between the microarrays with Goat Anti-mouse IgG of 0.02 mg/mL using the obliqueincidence reflectivity difference(OIRD)method.We obtained the reaction results and the reaction dynamic curves of 500 protein dots.In addition,we also used label-free detection of protein microarrays of 10080 sample dots,including BSA and different concentrations of Mouse IgG and Rabbit IgG,by OIRD.The obtained reaction results between the protein microarray with 1 mg/mL Goat Anti-mouse IgG and 1 mg/mL Goat Anti-rabbit IgG are reported herein.Experimental results show that OIRD can be not only label-free high-throughput detection method for biological microarrays but also label-free real-time detection in the interaction processes of biomolecules.
