酶联免疫吸附测定微孔板：与磁珠相比，它是用于蛋白质治疗药物免疫亲和液相色谱-串联质谱定量分析的可行免疫捕获平台吗？
ELISA microplate: a viable immunocapture platform over magnetic beads for immunoaffinity-LC-MS/MS quantitation of protein therapeutics?
作者信息
Yang Wenchu, Kernstock Robert, Simmons Neal, Alak Ala
机构信息
Bioanalysis-US, Astellas Research Institute of America, Skokie, IL 60077, USA.
出版信息
Bioanalysis. 2015;7(3):307-18. doi: 10.4155/bio.14.250.
AIM
Evaluate the performance of ELISA microplates versus commonly used magnetic beads for biological sample cleanup and/or enrichment in immunoaffinity-LC-MS/MS to reduce tedious beads washing procedures and a relatively high assay cost.
MATERIALS & METHODS: ELISA microplates were used as immunicapture platform and compared with magnetic beads for sample cleanup for LC-MS/MS quantitation of protein therapeutics.
RESULTS
One unmodified and two surface-activated microplates provided comparable linear ranges and sensitivities for a therapeutic protein (mass 78 kDa) using a human serum sample of 100 µl with 1:1 dilution compared with Tosylactivated magnetic beads using 200 µl of human serum without sample dilution. The assays' precision and accuracy were all within acceptable ranges. No nonspecific binding or other selectivity issues were observed.
CONCLUSION
The results suggested an ELISA microplate could be a viable immunocapture platform for immunoaffinity-LC-MS/MS quantitation of protein therapeutics.
目的
评估酶联免疫吸附测定（ELISA）微孔板与常用磁珠在免疫亲和液相色谱-串联质谱（immunoaffinity-LC-MS/MS）中用于生物样品净化和/或富集的性能，以减少繁琐的磁珠洗涤步骤和相对较高的检测成本。
材料与方法
将ELISA微孔板用作免疫捕获平台，并与磁珠进行比较，用于蛋白质治疗药物的LC-MS/MS定量分析中的样品净化。
结果
使用100µl 1:1稀释的人血清样品，一块未修饰的和两块表面活化的微孔板对一种治疗性蛋白质（质量78 kDa）提供了与使用200µl未稀释人血清的甲苯磺酰活化磁珠相当的线性范围和灵敏度。检测的精密度和准确度均在可接受范围内。未观察到非特异性结合或其他选择性问题。
结论
结果表明ELISA微孔板可能是用于蛋白质治疗药物免疫亲和LC-MS/MS定量分析的可行免疫捕获平台。
Zotero 插件