Recombinant human interferon a2b(rhIFNa2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNa2b is complex.In this study,an anti-rhI...Recombinant human interferon a2b(rhIFNa2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNa2b is complex.In this study,an anti-rhIFNa2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNa2b.RhIFNa2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNa2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNa2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNa2b detection assay had a limit of detection of 1 mg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other commercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.展开更多
目的:探讨雾化吸入不同剂量重组人干扰素α2b注射液对病毒性肺炎患儿血清炎性因子水平及免疫功能的影响。方法:选择2015年1月至2017年6月我院收治的病毒性肺炎患儿140例,按照给药剂量分为高剂量组(n=46)、低剂量组(n=48)和对照组(n=46)...目的:探讨雾化吸入不同剂量重组人干扰素α2b注射液对病毒性肺炎患儿血清炎性因子水平及免疫功能的影响。方法:选择2015年1月至2017年6月我院收治的病毒性肺炎患儿140例,按照给药剂量分为高剂量组(n=46)、低剂量组(n=48)和对照组(n=46)。三组患儿均接受病毒性肺炎常规治疗,高剂量组给予重组人干扰素α2b注射液每次20万U/kg雾化吸入治疗,低剂量组给予重组人干扰素α2b注射液每次10万U/kg雾化吸入治疗,对照组给予0.9%生理盐水2 m L雾化吸入治疗。观察三组患儿临床疗效、不良反应以及治疗前、治疗3周后血清炎症因子水平和免疫功能的变化。结果:高剂量组和低剂量组总有效率高于对照组(P<0.05),高剂量组和低剂量组总有效率比较差异无统计学意义(P>0.05)。三组患儿不良反应发生率比较差异无统计学意义(P>0.05)。治疗3周后三组患儿血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)水平均较治疗前降低(P<0.05),高剂量组IL-6、TNF-α、CRP水平低于低剂量组和对照组(P<0.05),低剂量组IL-6、TNF-α、CRP水平低于对照组(P<0.05)。治疗3周后高剂量组和低剂量组血清CD3^+、CD4^+、CD4^+/CD8^+较治疗前升高,CD8^+较治疗前降低(P<0.05);对照组CD3^+、CD4^+/CD8^+较治疗前显著升高,CD8^+较治疗前显著降低(P<0.05),CD4^+比较差异无统计学意义(P>0.05)。高剂量组CD3^+、CD4^+、CD4^+/CD8^+高于低剂量组和对照组,CD8^+低于低剂量组和对照组(P<0.05),低剂量组CD3^+、CD4^+、CD4^+/CD8^+高于对照组,CD8^+低于对照组(P<0.05)。结论:雾化吸入重组人干扰素α2b注射液可以降低病毒性肺炎患儿血清炎性因子水平,增强患儿细胞免疫功能,每次20万U/kg雾化吸入的效果优于10万U/kg,值得临床推广应用。展开更多
基金support was provided by the National Science and Technology Major Project(Grant No.:2015ZX09501008)。
文摘Recombinant human interferon a2b(rhIFNa2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNa2b is complex.In this study,an anti-rhIFNa2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNa2b.RhIFNa2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNa2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNa2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNa2b detection assay had a limit of detection of 1 mg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other commercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.
文摘目的:探讨雾化吸入不同剂量重组人干扰素α2b注射液对病毒性肺炎患儿血清炎性因子水平及免疫功能的影响。方法:选择2015年1月至2017年6月我院收治的病毒性肺炎患儿140例,按照给药剂量分为高剂量组(n=46)、低剂量组(n=48)和对照组(n=46)。三组患儿均接受病毒性肺炎常规治疗,高剂量组给予重组人干扰素α2b注射液每次20万U/kg雾化吸入治疗,低剂量组给予重组人干扰素α2b注射液每次10万U/kg雾化吸入治疗,对照组给予0.9%生理盐水2 m L雾化吸入治疗。观察三组患儿临床疗效、不良反应以及治疗前、治疗3周后血清炎症因子水平和免疫功能的变化。结果:高剂量组和低剂量组总有效率高于对照组(P<0.05),高剂量组和低剂量组总有效率比较差异无统计学意义(P>0.05)。三组患儿不良反应发生率比较差异无统计学意义(P>0.05)。治疗3周后三组患儿血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)水平均较治疗前降低(P<0.05),高剂量组IL-6、TNF-α、CRP水平低于低剂量组和对照组(P<0.05),低剂量组IL-6、TNF-α、CRP水平低于对照组(P<0.05)。治疗3周后高剂量组和低剂量组血清CD3^+、CD4^+、CD4^+/CD8^+较治疗前升高,CD8^+较治疗前降低(P<0.05);对照组CD3^+、CD4^+/CD8^+较治疗前显著升高,CD8^+较治疗前显著降低(P<0.05),CD4^+比较差异无统计学意义(P>0.05)。高剂量组CD3^+、CD4^+、CD4^+/CD8^+高于低剂量组和对照组,CD8^+低于低剂量组和对照组(P<0.05),低剂量组CD3^+、CD4^+、CD4^+/CD8^+高于对照组,CD8^+低于对照组(P<0.05)。结论:雾化吸入重组人干扰素α2b注射液可以降低病毒性肺炎患儿血清炎性因子水平,增强患儿细胞免疫功能,每次20万U/kg雾化吸入的效果优于10万U/kg,值得临床推广应用。
