RESUMO
Objective:To prepare a colloid gold kit for the qualitative detection of the dopes met-amphetamine and morphine by one-step chromatography immunoassay in human samples based on the principle of the highly specific immunochemical reactions between the antigens and their antibodies which were used for the analysis of specific compounds in human urine samples at ideal cut-off concentrations.Methods:The monoclonal antibodies(McAb) of either mouse anti-morphine or mouse met-amphetamine were prepared by hybridomas and the limiting dilution respectively. The titers of McAb were tested by ELISA.The kit was assembled and the specificity,sensitivity,and stability were investigated.In addition,these characters were compared with those of issued Pan Probe single test strip.Results:①The titer of the first McAb against morphine was 1∶3.2?103, as for another, 1∶1.6?103. ②The colloid gold kit was found with following cut-off for the dope concentrations,morphine at 300 ng/ml and met-amphetamine at 1 000 ng/ml. ③The colloid gold kit showed fine specificity without cross-reaction with routine medicines for cold treatment or with the solf-drinks of cokocola or with tobacco for the individuals who did not take the dopes the 7 days.④The colloid gold kit was even more sensitive than the issued strip,because urine sample of the junkies were still positive 4-5 days post-intake.Conclusion:The colloid gold kit for indentifying the drug-users is a rapid,specific, sensitive immunoassay suitable for the simultaneous, qualitative detection of drug abuse.
RESUMO
A sandwich ELISA for the measurement of urokinase(UK)antigen was developed based on anti—UK monoclonal antibodies(McAbs)against two non—overlapping epitopes.The lower limit of sensitivity of the assay was 0.15ng/ml.Coefficients of variation of the assay at physilogi-cal levels of UK were 4.3 percent within assaysw and 8.7 percent between assays.The recovery of added UK was about 98 percent.Culture media of some human malignant cell lines contains more UK than that of normal cell lines measured by our assay.The ELISA was used to measurethe concentration of UK in plasma from 82 healthy donors.The mean value for the healthy donors was 1.31?60.6ng/ml of UK in plasma.
RESUMO
Two kinds of mouse monoclonal antibodies which recognize distinct epitopes of the IL-2 receptor (a chain) were used to establish a sandwich ELISA for measuring soluble IL-2 receptors. The results indicated that this method is sensitive and specific for detecting soluble IL-2 receptors of cell culture supernatant and serum from both healthy donor and patients with leukemia. Therefore this sandwich ELISA method is useful for basic and clinic immunology research.
RESUMO
Using 22 monoclonal antibodies (McAbs) as probe to analyse the antigenic determinants of crystallized Tian Hua Fen (THF) or Trichosanthin protein in ELISA additivity test was reported. The 22 McAbs were come from five fusions, each from different fusion well. The isotype of 22 McAbs were identified by immunodiffusioo test, 20 McAbs belong to IgG_1 subclass, 1 IgG_(2a) subclass and 1 IgM class. Based on the results obtained from ELISA additivity test, 22 McAbs could be clustered into six distinct groups. It indicates that there are six different antigenic determinants in crystallized THF, each can be recognized by some McAbs. The results coincide with the calculation of B cell recognized antigenic determinants obtained by prediction from amino acid sequences of THF protein, The work has laid a foundation for further exploring the structure-function relationship of THF protein on molecular level .