کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2088483 | 1545743 | 2011 | 15 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
MALDI Immunoscreening (MiSCREEN): A method for selection of anti-peptide monoclonal antibodies for use in immunoproteomics
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کلمات کلیدی
mAbsSISCAPAMRMKLHMALDI-TOFPBS3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate - 3 - [(3-کلامیدوپروپیل) دی متیل آمونیو] -1-پروپان سولفوناتenzyme linked immunosorbent assay - آنزیم تست ایمونوسیورسانس مرتبط استHigh affinity - بالا بودن رابطهELISA - تست الیزاSPR - تشدید پلاسمون سطحیSurface plasmon resonance - تشدید پلاسمون سطحیequilibrium dissociation constant - تعادل تعادل ثابتassociation rate constant - ثابت نرخ ارتباطMass spectrometry - طیف سنجی جرمیPhosphate buffered saline - فسفات بافر شورMatrix-Assisted Laser Desorption/Ionization Time-of-Flight - مدت زمان پرواز یونیزاسیون لیزر ماتریکس کمک می کندmultiple reaction monitoring - نظارت چندگانه چندگانهMonoclonal antibodies - پادتنهای تَکتیرهCHAPS - چاپسhigh performance liquid chromatography - کروماتوگرافی مایع با کارایی بالاHPLC - کروماتوگرافی مایعی کاراkeyhole limpet hemocyanin - کلم هول limpet هموسیانین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: MALDI Immunoscreening (MiSCREEN): A method for selection of anti-peptide monoclonal antibodies for use in immunoproteomics MALDI Immunoscreening (MiSCREEN): A method for selection of anti-peptide monoclonal antibodies for use in immunoproteomics](/preview/png/2088483.png)
چکیده انگلیسی
A scalable method for screening and selection of peptide-specific monoclonal antibodies (mAbs) is described. To identify high affinity anti-peptide mAbs in hybridoma supernatants, antibodies were captured by magnetic affinity beads followed by binding of specific peptides from solution. After timed washing steps, the remaining bound peptides were eluted from the beads and detected by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). This allowed measurement of monovalent interactions of peptides with single antigen binding sites on the antibodies, thus reflecting antibody affinity rather than avidity. Antibodies that were able to bind target peptides from solution phase and retain them during washing for a minimum of 10Â min were identified by the strength of the appropriate m/z peptide MS signals obtained. This wash time reflects the minimum peptide dissociation time required for use of these antibodies in several current immuno-mass spectrometry assays. Kinetic analysis of antibody-peptide binding by surface plasmon resonance (SPR) showed that the selected antibodies were of high affinity and, most importantly, had low dissociation constants. This method, called MALDI immunoscreening (MiSCREEN), thus enables rapid screening and selection of high affinity anti-peptide antibodies that are useful for a variety of immunoproteomics applications. To demonstrate their functional utility in immuno-mass spectrometry assays, we used the selected, purified RabMAbs to enrich natural (tryptic) peptides from digested human plasma.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 364, Issues 1â2, 1 February 2011, Pages 50-64
Journal: Journal of Immunological Methods - Volume 364, Issues 1â2, 1 February 2011, Pages 50-64
نویسندگان
Morteza Razavi, Matthew E. Pope, Martin V. Soste, Brett A. Eyford, Angela M. Jackson, N. Leigh Anderson, Terry W. Pearson,