ABSTRACT
Eight mouse hybridoma cell lines which stably secreted monoclonal antibodies ( McAbs ) against human prostate-specific antigen-α1-antichymotrypsin complex ( PSA-ACT ) were obtained through hybridoma technique. After purification, the immunological characters of 8 McAbs were identified and classified by epitopes analysis through indirect enzyme-linked immunosorbent assay ( ELISA) . A pair of McAbs was chosen from above 8 McAbs, based on which a highly sensitive, simple and rapid chemiluminescence enzyme immunoassay ( CLEIA) was developed for determination of PSA-ACT in human serums using the lumino-H2 O2 reaction catalyzed by horseradish peroxidase ( HRP) as the chemiluminescence system. Several experiment factors such as coating buffer, coating concentration, dilution ratio of PSA-ACT-HRP complex, incubation time, immunoreaction protocol and chemiluminescence reaction time were optimized. The results showed that the linear range of the proposed method for PSA-ACT determination was 0-40 ng/mL (R2=0. 9943), with the detection limit of 0. 53 ng/mL. The inter-assay relative standard deviations (RSDs) were 4. 6%-6. 6%, and intra-assay RSDs were 5 . 7%-8 . 0%. The recoveries of PSA-ACT at three spiked levels in serum samples were 95. 4%-104. 2%. The proposed method exhibited a cross-reactivity of 0. 6% with free-PSA. The proposed method is stable, sensitive, rapid and simple, and provides a foundation for the development of PSA-ACT CLEIA kit and shows great value in clinical auxiliary diagnosis of prostate cancer.
ABSTRACT
Objective To investigate the relationship between serum hepatitis B virus markers (HBV-M ) and HBV-DNA . Methods Enhanced chemiluminescence enzyme immunoassay (ECLIA ) and real-time fluorescent quantitative polymerase chain re-action(FQ-PCR) were employed to detect HBV-M and HBV-DNA in 262 serum samples ,respectively .HBV surface antigen(HB-sAg) ,anti-HBV surface antibody(HBsAb) ,HBV e antigen(HBeAg) ,anti-HBV e antibody(HBeAb) ,anti-HBV core antibody(HB-cAb) were included in HBV-M .Results Compared the positive rates of HBV-DNA ,HBsAg ,HBeAb in HBeAg-negative patients with those in HBeAg-positive patients ,the differences were statistically significant (P< 0 .01) .29(36 .7% ) patients with HBV-DNA logarithm value not less than 5 were found in HBeAg-negative patients .Differences of HBeAg ,HBeAb positive rates among patients with different ages were statistically significant (P<0 .01) .25 patients with HBsAg less than 250 IU/mL were found in HBV-DNA-positive patients ,12(48 .0% ) of which showed HBV-DNA logarithm value not less than 5 .HBV-DNA logarithm value of HBV-DNA-positive patients was positively correlated to HBeAg and HBeAb (r= 0 .542 ,0 .607 ,P< 0 .01) .Conclusion Com-bined detection of HBV-M and HBV-DNA contributes to estimating the HBV infection .
ABSTRACT
A chemiluminescence enzyme immunoassay based on magnetic microparicles (MmPsCLEIA) was developed to evaluate serum α-fetoprotein (AFP) in parallel with tramional colorimetric enzyme-linked immunsorbrnt assay (ELISA).A sestematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer of immunoreagents,less total assay time,and better linearity,recovery,precision,senitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results werecompared with commercial electrochemilunminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R2=0.6703; however,the correlation between MPs-CLEIA and ECLIA (R2=0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R2=0.6866).
ABSTRACT
Glypican-3 (GPC3) is reported as a great promising tumor marker for hepatocellular carcinoma (HCC) diagnosis.Highly sensitive and accurate analysis of serum GPC3 (sGPC3),in combination with or instead of traditional HCC marker alpha-fetoprotein (AFP),is essential for early diagnosis of HCC.Biomaterial-functionalized magnetic particles have been utilized as solid supports with good biological compatibility for sensitive immunoassay.Here,the magnetic nanoparticles (MnPs) and magnetic microparticles (MmPs) with carboxyl groups were further modified with streptavidin,and applied for the development of chemiluminescence enzyme immunoassay (CLEIA).After comparing between MnPs- and MmPs-based CLEIA,MnPs-based CLEIA was proved to be a better method with less assay time,greater sensitivity,better linearity and longer chemiluminescence platform.MnPs-based CLEIA was applied for detection of sGPC3 in normal liver,hepatoeirrhosis,secondary liver cancer and HCC serum samples.The results indicated that sGPC3 was effective in diagnosis of HCC with high performance.