基于报告基因的抗IFNR1单抗生物学活性测定方法的建立

目的：建立抗Ⅰ型干扰素受体亚基1单抗（抗IFNR1单抗）的生物学活性检测方法。方法：利用HEK293-ISRE-Luc细胞系，通过荧光素酶检测系统（ONE-Glo^® Luciferase Assay System），进行抗IFNR1单抗生物学活性检测，并对该方法进行精密度、准确度、专属性的验证。结果：抗IFNR1单抗在该方法中均存在量效关系，利用四参数方程拟合成4S型曲线，50%、75%、100%、125%及150%的加标样品相对效价（n=3）分别为（49.58±2.90）%（RSD=5.8%）、（76.27±5.12）%（RSD=6.7%）、（99.84±4.21）%（RSD=4.2%）、（123.08±2.58）%（RSD=2.1%）、（158.69±4.72）%（RSD=3.0%），RSD均小于10%；加样回收率分别为（99.16±5.81）%、（101.69±6.82）%、（99.84±4.21）%、（98.47±2.06）%和（105.80±3.14）%，RSD均小于10%；专属性良好。结论：建立的抗IFNR1单抗生物学活性检测方法，可作为抗IFNR1单抗生物学活性的常规检测方法。

Objective:To establish a biological activity assay for anti-type Ⅰ interferon receptor subunit 1 monoclonal antibody (anti-IFNR1 monoclonal antibody).Methods:The bioassay of anti-IFNR1 antibody was tested by cell line of HEK293-ISRE-Luc and ONE-Glo^® Luciferase Assay System, and the precision, accuracy and specificity of the method were validated.Results:The anti-IFNR1 antibody has dose-effect relation which could be fitted by four-parameter curve.The relative potencies of 50%, 75%, 100%, 125% and 150% of sample concentration(n=3) were(49.58±2.90)%, (76.27±5.12)%, (99.84±4.21)%, (123.08±2.58)%, (158.69±4.72)%, and RSDs were 5.8%, 6.7%, 4.2%, 2.1%, 3.0%, respectively.The RSD was less than 10%.The recoveries(n=3) were(99.16±5.81)%, (101.69±6.82)%, (99.84±4.21)%, (98.47±2.06)% and(105.80±3.14)%, respectively.RSDs were less than 10%.Specificity of the method was fine.Conclusion:The bioassay method of anti-IFNR1 antibody has been established, which can be a common bioassay method of anti-IFNR1 monoclonal antibody.

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