生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2013年
3期
366-369
,共4页
李丽%刘运龙%陈知航%张玉民%刘学龙%程远国
李麗%劉運龍%陳知航%張玉民%劉學龍%程遠國
리려%류운룡%진지항%장옥민%류학룡%정원국
重组人血清白蛋白%抗体检测%桥连ELISA%免疫清除
重組人血清白蛋白%抗體檢測%橋連ELISA%免疫清除
중조인혈청백단백%항체검측%교련ELISA%면역청제
recombinant human serum albumin%antibody assay%bridging ELISA%immune clearance
目的:建立一种高灵敏度、高特异性、操作简单快捷、通量高的重组人血清白蛋白(rHSA)抗体检测方法。方法:采用桥连ELISA法,即将rHSA包被于96孔板,加入待测血样及阳性对照,用辣根过氧化物酶标记的rHSA检测,显色读取D450nm/D570nm值;用此方法确定临界值、方法灵敏度、精密度、血药浓度对检测方法的影响,再以免疫清除法进行确证。结果:通过桥连ELISA法确定临界值为0.0492,方法灵敏度为352 ng/mL,方法板间、板内精密度均小于20%,且血药中的rHSA浓度为20μg/mL时不影响抗体的检测;经免疫清除法可将假阳性样本排除,从而提高了方法的特异性。结论:建立的方法可以准确、快速地检测出rHSA的特异性抗体。
目的:建立一種高靈敏度、高特異性、操作簡單快捷、通量高的重組人血清白蛋白(rHSA)抗體檢測方法。方法:採用橋連ELISA法,即將rHSA包被于96孔闆,加入待測血樣及暘性對照,用辣根過氧化物酶標記的rHSA檢測,顯色讀取D450nm/D570nm值;用此方法確定臨界值、方法靈敏度、精密度、血藥濃度對檢測方法的影響,再以免疫清除法進行確證。結果:通過橋連ELISA法確定臨界值為0.0492,方法靈敏度為352 ng/mL,方法闆間、闆內精密度均小于20%,且血藥中的rHSA濃度為20μg/mL時不影響抗體的檢測;經免疫清除法可將假暘性樣本排除,從而提高瞭方法的特異性。結論:建立的方法可以準確、快速地檢測齣rHSA的特異性抗體。
목적:건립일충고령민도、고특이성、조작간단쾌첩、통량고적중조인혈청백단백(rHSA)항체검측방법。방법:채용교련ELISA법,즉장rHSA포피우96공판,가입대측혈양급양성대조,용랄근과양화물매표기적rHSA검측,현색독취D450nm/D570nm치;용차방법학정림계치、방법령민도、정밀도、혈약농도대검측방법적영향,재이면역청제법진행학증。결과:통과교련ELISA법학정림계치위0.0492,방법령민도위352 ng/mL,방법판간、판내정밀도균소우20%,차혈약중적rHSA농도위20μg/mL시불영향항체적검측;경면역청제법가장가양성양본배제,종이제고료방법적특이성。결론:건립적방법가이준학、쾌속지검측출rHSA적특이성항체。
Objective: To develop a high specific, high sensitive bridging ELISA method for the quantification of recombinant human serum albumin(rHSA) antibody. Methods: The microplate was coated with rHSA, and a HRP-labeled rHSA was used as the detection antibody for the bridging ELISA. The established bridging ELISA would be evaluated by the cut off, sensitivity, precision, plasma concentration to the influence of method. Use im?mune clearance to prove the method convincingly. Results: The cut off value was 0.0492, the sensitivity of the bridging ELISA with a limit of detection was 352 ng/mL, the intra- and inter-assay precisions were less than 20%, and the plasma concentration was 20 μg/mL that didn't influence the detection of antibody. The immune clearance could exclude false positive samples. Conclusion: This method is accurate, fast for the determination of rHSA antibody.