کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902385 | 1545802 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Use of biotinylated 17β-estradiol in enzyme-immunoassay development: Spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin-antibody binding
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کلمات کلیدی
ACNN-(3-dimethylaminopropyl)-N′-ethylcarbodiimideBMETMBRIAEDCIgG17β-estradiol - 17β استرادیول2-(N-morpholino)ethanesulfonic acid - 2- (N-مورفولینو) اتان سولفونیک اسید3,3′,5,5′-tetramethylbenzidine - 3،3 '، 5،5'-تترامیلیل بنزیدینBSA - BSADMSO - DMSORadio immunoassay - آزمایش ایمنی رادیوییbovine serum albumin - آلبومین سرم گاوAcetonitrile - استونیتریلenzyme immunoassay - ایمونواسی آنزیم immunoglobulin G - ایمونوگلوبولین GEIA - اینELISA - تست الیزاAbsorbance - جذبTemperature - دماRoom temperature - دمای اتاقDimethylsulfoxide - دیمتیل سولفواکسیدSpacer length - طول فاصلهMass spectrometry - طیف سنجی جرمیMeS - مسAntibody - پادتَن یا آنتیبادیhigh performance liquid chromatography - کروماتوگرافی مایع با کارایی بالاHPLC - کروماتوگرافی مایعی کارا
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Use of biotinylated 17β-estradiol in enzyme-immunoassay development: Spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin-antibody binding Use of biotinylated 17β-estradiol in enzyme-immunoassay development: Spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin-antibody binding](/preview/png/9902385.png)
چکیده انگلیسی
17β-estradiol (E2) concentrations are in the low pg/ml range in plasma. To develop a sensitive enzyme immunoassay (EIA) for E2-determination a highly specific antibody raised against a 6-carboxymethyl (CMO)-E2-bovine serum albumine conjugate was used. Based on 6-CMO-E2 and 6-amino-E2, four biotinylated tracers with two different spacer lengths between E2 and biotin were synthesized using biotinylation reagents in one step reactions. All amino-based tracers were unsuitable for assay development because the antibody binding was too weak compared to the analyte E2. For 6-CMO-based tracers the simultaneous binding of the tracer to the antibody and streptavidin seems to be the determining step in the procedure depending on incubation temperature and spacer lengths. While a short spacer of 9 carbon atoms was susceptible to room temperature, a longer spacer of 16 carbon atoms showed nearly the same results for incubation at 4 °C or at room temperature. The absolute detection limit of this system was 0.63 pg/well. For sample clean-up, porcine plasma was solvent-extracted and depending on the initial plasma volume further purified by solvent partition. Determination of reproducibility resulted in intraassay coefficients of variation of 13% and 5.3% for samples with E2-levels of 15 pg/ml and 236 pg/ml, respectively. Measurement of E2-spiked blood plasma revealed recoveries of 83% up to 100% for E2 concentrations between 50 pg/ml and 1000 pg/ml. Only for the lowest concentration (20 pg/ml) a recovery of 58% was observed. Correlation of the EIA with an established radio immunoassay resulted in r=0.991 using the same antibody.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 297, Issues 1â2, February 2005, Pages 225-236
Journal: Journal of Immunological Methods - Volume 297, Issues 1â2, February 2005, Pages 225-236
نویسندگان
Markus Lacorn, Kerstin Fleischer, Stephanie Willig, Sabine Gremmel, Hans Steinhart, Rolf Claus,