| Objective: To obtain and identify the monoclone antibodies of 8-iso-prostaglandin F 2alpha (8-iso-PGF2α) ; to manufacture the determining kit of 8-iso-PGF2α (ELISA); to determinate the contents of urine 8-iso-PGF2 α in normal neonates and neonates with hypoxic-ischemic encephalopathy (HIE). Methods: Hapten and 8-iso-PGF2α is conjugated with protein BSA/OVA by the methods of EDC. The spleen cells from BALB/C mice immunized with GTX2/3-BSA are fused with murine Sp2/0 myeloma cells. The specific hybridoma cells and the ascites were analyzed using ELISA. Ascites produced by positive clone is purified. HRP was used to marked antigen to manufacture the determining kit of 8-iso-PGF2α (ELISA). Urine samples from normal (126 cases) and HIE neonates (151 cases) were collected. EIA assay was used to determine urine 8-iso-PGF2a content. Results: One strain of cells has been obtained, which can secrete highly specific monoclonal antibodies against 8-iso-PGF2α . The coated concentration of anti-8-iso-PGF2α was 1μ g/ml, incubation time last 30min, the cutoff value was 0.140+NC, 100μl samples is necessary for 8-iso-PGF2α assasy, and there was no change in OD450 value when the kits were placed for 7days at 37℃. For urine 8-iso-PGF2a content, normal neonates were 27.4 ± 9.8 ng/mmol·Cr; light, moderate, serious HIE neonates were 65.3±13.2, 154.6±31.6 and 241.7±41.0 ng/mmol·Cr, respectively (p<0.01) . Using 47.0, 91.7 and 217.5 ng/mmol·Cr of urine 8-iso-PGF2α as the boundary of distinguishing between normal, mild, moderate and severe HIE neonates, the sensitivity and specificity were 95.2% and 99. 2%, 100% and 95. 2%, 65. 8% and 100%, respectively.Conclusion: Monoclone antibodies of 8-iso-PGF2α can be obtained by means of antibody engineering technique. 8-iso-PGF2α kit (ELISA) has a good stabiliza-tivity, and can accurately determine urine 8-iso-PGF2α contents in neonates. The determination of urine 8-iso-PGF2 α may be a reliable and simple biochemical index to reflect the condition of HIE.
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