Determination of MIL50, a humanized antibody, in different tissues of rats by a sandwich enzyme-linked immunosorbent assay

VernacularTitle:双夹心酶联免疫法检测生物样品中的人源化抗体MIL50
Author: Yifan WU ; Peiyuan JIA ; Junhua WU ; Jianping ZHOU ; Haixia LI
Publication Type:Journal Article
Keywords: ricin; humanized antibody; enzyme-linked immunosorbent assay
From: Military Medical Sciences 2013;(11):827-830
CountryChina
Language:Chinese
Abstract: Objective To develop a sandwich enzyme-linked immunosorbent assay ( ELISA) for the determination of a humanized antibody MIL50.Methods RiVax, a mutant of RTA (a chain of ricin) expressed in E.coli, was used as the coating protein.Horseradish peroxidase (HRP) labeled IgG of goats against humans was used to determine MIL 50 captured by RiVax coated on the plate.Results Compared with ricin, RiVax could bind well with MIL50,indicating it could be used as a coating protein to determine MIL50 instead of holotoxin.Using this method, the detective limit of MIL50 in PBST (PBS containing 0.1%Tween 20) could be as low as 0.030 mg/L.The absorbance value of ELISA for MIL50 in the ser-um and homogenate of the liver , spleen, lung, kidney, muscle of rats was lower than that in PBST .Conclusion The sandwich ELISA is a sensitive method for the analysis of distribution of MIL 50 in rat tissues.