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Immunoassay Method (immunoassay + method)

Distribution by Scientific Domains
Medical Sciences71%
Life Sciences28%

Kinds of Immunoassay Method

  • enzyme immunoassay method
    		•

    Selected Abstracts

    Construction of an antibody microarray based on agarose-coated slides

    ELECTROPHORESIS, Issue 3 2007
    Lin-Li Lv
    Abstract The antibody microarray, a high-throughput multiplex immunoassay method, has become a significant tool for quantitative proteomics studies. We describe here the strategies for optimizing the condition of antibody microarray building based on agarose-coated slides. In this study, modified glass slides were robotically printed with capture antibodies against monocyte chemoattractant protein 1 (MCP-1), then dilutions of the cytokine were applied to the arrays, and the protein was detected with biotin-labeled antibody coupled with Cy3-conjugated streptavidin. Thus a protein profiling microarray based on sandwich immunoassay has been established. Various factors in the production of antibody microarrays were analyzed: the capture antibody concentrations, shelf life of the postprinting slides, blocking buffers, and reproducibility of the system. A calibration curve with a correlation coefficient of 0.9995 was established which suggested that the matrix can retain arrayed proteins in near-quantitative fashion. The results revealed high signal uniformity and reproducibility with regard to intra-array (1.3%) and the interarray (8.7%) variation at the capture antibody concentration of 125,µg/mL. Besides, the printed arrays could be stored for at least two months without any apparent change of the performance parameters. [source]

    Interference in thyroid-stimulating hormone determination

    EUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 8 2010
    Mauro Imperiali
    Eur J Clin Invest 2010; 40 (8): 756,758 Abstract Background, Thyroid-stimulating hormone (TSH) measurement plays a major role in the diagnosis of thyroid disorders. Despite the good quality of immunochemical tests measuring TSH levels, the presence of interfering substances can sometimes alter the TSH results. Design, We reported the case of a 79-year-old man affected by primary autoimmune hypothyroidism hospitalized for pneumonia. A TSH value > 100 mIU L -1 (reference: 0.44 mIU L -1) was found at admission. No signs and symptoms of hypothyroidism were found upon clinical examination and serum concentration of the free thyroxine (FT4) was normal. Results, Serum treatment in heterophile antibody blocking tubes did not change the TSH result in our assay, while normal levels were found in a different immunoassay method. An abnormal pattern was found in protein electrophoresis at admission, with IgG / j and IgM / k monoclonal bands proved in immunofixation. Interestingly, the disappearance of monoclonal bands was paralleled with a normalization of the TSH value. Conclusions, We suggest in this study that the TSH determination might be influenced by the presence of transient paraproteins. [source]

    Investigation of the relationship between low Apgar scores and early neonatal thyroid function

    PEDIATRICS INTERNATIONAL, Issue 5 2000
    Erbil Sak
    Abstract Background: The aim of the present study was to assess the effects of low Apgar scores on perinatal thyroid function. Methods: Forty full-term infants delivered by the normal spontaneous vaginal route were enrolled into the study. All babies had 1 and 5 min Apgar scores below 4. The control group consisted of 26 full-term healthy neonates. Cord blood and serum tri-iodothyronine (T3), thyroxine (T4), reverse tri-iodothyronine (rT3), free thyroxine (FT4), thyroid-stimulating hormone (TSH) and thyroid-binding globulin (TBG) determinations were performed by an enzyme immunoassay method. Results: The mean values of FT4 and T4 observed in the cord blood of the study group were significantly lower compared with matched controls, whereas the mean TSH values were significantly higher. There were no differences in concentrations of T3, rT3 and TBG between the two groups. Conclusions: These results demonstrate the existence of transient hypothyroidism at birth in babies with Apgar scores below 4 delivered by the spontaneous vaginal route. [source]

    Cloning, expression, and identification of anti-carbofuran single chain Fv gene

    BIOTECHNOLOGY PROGRESS, Issue 4 2009
    Hong Wang
    Abstract Phage display method was used to clone anti-carbofuran (CBF) single chain Fv (scFv) gene. The heavy chain and light chain variable region genes were amplified by the polymerase chain reaction from the CBF-specific hybridoma cell lines 5D3 and assembled as a scFv DNA fragment with linker peptide (Gly4Ser)3. The scFv DNA fragment was cloned into M13 phagemid vector pCANTAB5E and the anti-CBF antibody libraries were then constructed. After one round of panning with CBF-ovalbumin (CBF-OVA) as a conjugate, antigen-binding positive recombinant phage clones were successfully selected by enzyme-linked immunosorbent assay (ELISA). The positive phages were used to infect Escherichia coli HB2151 cells and the expression of the soluble scFv antibodies was then induced by IPTG. The scFv antibody was about 31 kDa by SDS-PAGE and showed HRP-anti-E-tag antibody-recognized activity by Western blotting. The indirect competitive ELISA (icELISA) showed that the recombinant scFv antibody could competitively combine with CBF, with the IC50 value of 1.07 ng/mL. The cross reactivity studies showed that the anti-CBF scFv antibody, similar to the parent monoclonal antibody, poses high specificity to CBF and has little reactivity to the analogs. Taken together, these findings suggest that the recombinant scFv antibody can be used for further developing immunoassay method for CBF. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source]

    Determination of medullasin levels for the diagnosis of multiple sclerosis

    ACTA NEUROLOGICA SCANDINAVICA, Issue 4 2000
    Y. Aoki
    Objectives , To obtain a simple and reliable clinical parameter for the diagnosis of multiple sclerosis among patients with neurological diseases. Patients and methods, Heparinized peripheral blood was obtained from patients with multiple sclerosis and those with non-inflammatory neurological diseases and healthy volunteers. A new enzyme immunoassay method determining medullasin levels in human granulocytes was developed by using mouse monoclonal antibody against medullasin. Results, A newly developed enzyme immunoassay method for medullasin can detect as little as 1 ng/ml medullasin and results can be obtained within 2 h. Eighty-five out of 112 patients with multiple sclerosis (75.8%) showed positive results (above means of normals+2 SD) in the medullasin test, while 15.4% (12/78) of patients with non-inflammatory neurological disease had positive results. Conclusion, This newly developed enzyme immunoassay method for medullasin is considered to be a useful paraclinical test for the diagnosis of multiple sclerosis. [source]

    Early iron supplementation in very low birth weight infants , a randomized controlled trial

    ACTA PAEDIATRICA, Issue 6 2009
    Mari Jeeva Sankar
    Abstract Aim: To evaluate if supplementing iron at 2 weeks of age improves serum ferritin and/or haematological parameters at 2 months of life in very low birth weight (VLBW) infants. Methods: Preterm VLBW infants who received at least 100 mL/kg/day of oral feeds by day 14 of life were randomized to either ,early iron' (3,4 mg/kg/day orally from 2 weeks) or ,control' (no iron until 60 days) groups. Infants were followed up fortnightly and all morbidities were prospectively recorded. Serum ferritin was measured at 60 days by enzyme immunoassay method. Results: Forty-six infants were included in the study; primary outcome was available for 42 infants. There was no difference in either serum ferritin (mean: 50.8 vs. 45.3 ,g/L; adjusted difference in means: 5.8, 95% CI: ,3.0, 14.6; p = 0.19) or haematocrit (32.5 ± 5.3 vs. 30.8 ± 6.3%; p = 0.35) at 60 days between the early iron and control groups. The magnitude of fall in serum ferritin from baseline to the end of study period was also not different between the groups (4.9 vs. 13.8 ,g/L; difference in means: 8.8; 95% CI: ,0.3, 17.9; p = 0.06). The requirement of blood transfusions (9.5 vs. 13%; p = 0.63) and a composite outcome of common neonatal morbidities (19% vs. 21.7%; p = 0.55) were also not different between the two groups. Conclusion: Supplementing iron at 2 weeks of age in preterm VLBW infants did not improve either serum ferritin or the haematological parameters at 2 months when compared to the standard practice of starting iron from 8 weeks of age. [source]

    Homocysteine levels in patients with risk factors for atherosclerosis

    CLINICAL CARDIOLOGY, Issue 6 2001
    Arnon Blum M.D.
    Abstract Background: Abundant epidemiological evidence has demonstrated that the presence of mild to moderate hyperhomocysteinemia is an independent risk factor for atherosclerosis in the coronary, cerebral, and peripheral vasculature, and for vascular disease, including coronary disease. It has been demonstrated that plasma total homocysteine level is a strong predictor of mortality in patients with angiographically confirmed coronary artery disease. Hypothesis: The study was undertaken to determine the extent of homocysteine levels in patients without documented coronary artery disease, but with at least one risk factor for atherosclerosis. Methods: Fasting blood samples were collected prospectively from 160 consecutive patients (50 women and 110 men, mean age 65 ± 7 years) who had at least one risk factor for atherosclerosis, but had no documented coronary artery disease. Homocysteine levels were measured by an immunoassay method. Results: Of the patients studied, 78 (48.75%) with at least one risk factor for atherosclerosis had high homocysteine levels; 62 patients had mild hyperhomocysteinemia (15,30 ,mol/l); and 16 patients had moderate hyperhomocysteinemia (30,100 ,mol/l). Conclusions: Our data suggest that hyperhomocysteinemia is highly prevalent in patients with risk factors for atherosclerosis. Homocysteine level (an independent convertible risk factor to atherosclerosis) should be measured routinely in patients with risk factors for atherosclerosis and treated appropriately. [source]