2種の異種抗体を用いたELISAによるα₂-アンチプラスミン抗原量の測定

Abstract

A new solid enzyme-linked immunosorbent assay (ELISA) for human α₂-antiplasmin (α₂-AP) antigen was developed using two kinds of anti-human α₂-AP antiserum. The procedure of α₂-AP antigen assay is as follows : Step 1 ; a microtiter plate is coated with an anti-human α₂-AP guineapig antiserum for 24 hrs at 4℃. Step 2 ; serially diluted plasma is added to the plate and incubated for 24 hrs at 4℃. Step 3 ; after washing with a phosphate buffered saline, an anti-human α₂-AP rabbit antiserum is added to the plate and incubated for 3 hrs at 37℃. Then, a peroxidase conjugated anti-rabbit IgG goat antiserum is added and incubated for 1 hr at 37℃. After the 3rd step, the microtiter plate is washed extensively. Finally, the α₂-AP antigen is determined by ELISA method using o-phenylendiamin as the substrate. The lower limit of α₂-AP antigen in this assay was 0.1 U/dl. In 25 healthy adult males and 25 adult females, the mean values of α₂-AP were 96.8±12.2 U/dl and 95.2±10.7 U/dl, respectively. A good correlation was found between the α₂-AP level determined by this ELISA method and by Laurell's method. There was also a good correlation between α₂-AP activity by an amidolytic method and antigen level by the ELISA.