Fluorimetric enzyme immunoassay for serum β2-microglobulin based on the horseradish peroxidase-labeled Fab′ fragment of immunoglobulin
✍ Scribed by Kenji Sugimoto; Kiyoshi Zaitsu; Yosuke Ohkura
- Publisher
- Elsevier Science
- Year
- 1985
- Tongue
- English
- Weight
- 475 KB
- Volume
- 169
- Category
- Article
- ISSN
- 0003-2670
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✦ Synopsis
A sensitive sandwich-type enzyme immunoassay for human p,-microglobulin in serum is described. Rabbit anti-human &-microglobulin IgG is coated on a styrene-maleic anhydride copolymer bead, which is incubated with p,-microglobulin in serum sample at 30°C for 1 h, and again incubated at 30°C for 1 h with a conjugate of anti-p,-microglobulin Fab' (a fragment of rabbit anti-human P,-microglobulin IgG) and horseradish peroxidase to form a sandwich-type immunocomplex. The conjugate is prepared with a heterobifunctional reagent, N-succinimidyl-3-(2-pyridy1dithio)propionat.e.
The peroxidase activity of the immunocomplex is measured spectrofluorimetrically by use of 3-@-hydroxyphenyl)propionic acid. The method permits precise assay of 0.05-10 ng of p,-microglobulin in serum, with calibration linearity up to 1.0 ng of protein.