1.Preparation and evaluation of a time-resolved fluoroimmunoassay kit for CA50 determination.
Jian-feng HANG ; Ying-song WU ; Wei-wen XU ; Wei-hong YU ; Ying HUANG ; Ming LI
Chinese Journal of Oncology 2007;29(3):189-192
OBJECTIVETo prepare a rapid and sensitive diagnostic kit for detection of CA50 based on time-resolved fluoroimmunoassay.
METHODSA sandwich-TRFIA diagmostic kit was developed using anti-CA50 monoclonal antibody and all parameters of the kit were evaluated.
RESULTSThe linear measurement range of the kit was (5 - 300) U/ml. The sensitivity was 0.2 U/ml. The intra- and inter-assay coefficients of variation were 4.3% - 8.2% and 7.7% - 11.2%, respectively. There was no cross-reaction with CEA, CA12-5, CA15-3 and AFP. The cross reactivity with CA19-9 was 0.7 U/ml. The correlation coefficient of detection results of 107 blood samples between this newly developed kit and commercially available CA50 RIA kit was 0.901.
CONCLUSIONThis newly developed CA50-TRFIA kit is a valuable test tool for clinical application with even better sensitivity, specificity and accuracy.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, Tumor-Associated, Carbohydrate ; blood ; Female ; Fluoroimmunoassay ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Radioimmunoassay ; Reagent Kits, Diagnostic ; standards ; Reference Standards ; Reproducibility of Results
2.The Effects of Anti-insulin Antibodies and Cross-reactivity with Human Recombinant Insulin Analogues in the E170 Insulin Immunometric Assay.
Serim KIM ; Yeo Min YUN ; Mina HUR ; Hee Won MOON ; Jin Q KIM
The Korean Journal of Laboratory Medicine 2011;31(1):22-29
BACKGROUND: Insulin assays are affected by varying degrees of interference from anti-insulin antibodies (IAs) and by cross-reactivity with recombinant insulin analogues. We evaluated the usefulness of the E170 insulin assay by assessing IA effects and cross-reactivity with 2 analogues. METHODS: Sera were obtained from 59 type 2 diabetes patients receiving continuous subcutaneous insulin infusion and 18 healthy controls. Insulin levels were determined using an E170 analyzer. To investigate the effects of IAs, we performed IA radioimmunoassays, and analyzed the differences between directly measured insulin (direct insulin) and polyethylene glycol (PEG)-treated insulins (free, IA-unbound; total, IA-bound and unbound insulin). We performed in-vitro cross-reactivity tests with insulin aspart and insulin glulisine. RESULTS: In IA-positive patients, E170 free insulin levels measured using the E170 analyzer were significantly lower than the direct insulin levels. The mean value of the direct/free insulin ratio and IA-bound insulin, which were calculated as the difference between total and free insulin, increased significantly as endogenous IA levels increased. The E170 insulin assay showed low cross-reactivities with both analogues (< 0.7%). CONCLUSIONS: IAs interfered with E170 insulin assay, and the extent of interference correlated with the IA levels, which may be attributable to the increase in IA-bound insulin, and not to an error in the assay. The E170 insulin assay may measure only endogenous insulin since cross-reactivity is low. Our results suggest that the measurement of free insulin after PEG pre-treatment could be useful for beta cell function assessment in diabetic patients undergoing insulin therapy.
Adult
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Aged
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Aged, 80 and over
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Cross Reactions
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Diabetes Mellitus, Type 2/blood/immunology
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Female
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Humans
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Infusions, Subcutaneous
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Insulin/analogs & derivatives/*blood/chemistry/immunology
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Insulin Antibodies/*blood
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Male
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Middle Aged
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Polyethylene Glycols/chemistry
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Radioimmunoassay/instrumentation/*methods
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Recombinant Proteins/analysis/immunology/metabolism