| Autor: |
Zben-Quan Guo, Yuanzong Li, Yong-Cheng Wang, Jin-Fu Zhao, Wen-Bao Chang |
| Rok vydání: |
2010 |
| Předmět: |
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| Zdroj: |
Chinese Journal of Chemistry. 19:1205-1210 |
| ISSN: |
1001-604X |
| DOI: |
10.1002/cjoc.20010191208 |
| Popis: |
A competitive indirect fluoroimmunoassay of free estradiol (E2) was established based on the thermal sensitivity of hydrogel–-poly-N-isopropylacrylamide. Free estradiol was covalently bound to bovine serum albumin (BSA) to form complete antigen (E2-BSA), which was in turn labeled by fluorescein isothiocyanate (FTTC) as the fluorescence probe. The anti- E2 monoclonal antibody (McAb) was prepared by an in vivo method, and coupled with N-isopropylacrylamide (NIPA) to make an immune copolymer, poly-N-isopropylacylamidemonoclonal antibody (pNIPA-McAb), for the determination of free E2. The immunoassay method was based on the competitive binding of free E2 and fluoresceinated antigen (E2-BSA-FTTC) with limited amount of pNIPA-McAb. When the immunological reaction was over, precipitation and centrifugal procedures were carried out to separate pNIPA-McAb-E2-BSA-FTTC from other constituents in solution. The precipitate pNIPA-McAb-E2-BSA-FTTC was dissolved in solution and then the fluorescence intensity was measured. The calibration curve covered a range of 78–500 ng/mL for free E2. The recoveries were 91.2–107.2%. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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