کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5522044 1545667 2017 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Research paperLabel free checkerboard assay to determine overlapping epitopes of Ebola virus VP-40 antibodies using surface plasmon resonance
ترجمه فارسی عنوان
آزمایش اکسپلویت رایگان برای تعیین اپیتوپهای همپوشانی آنتی بادی ویروس ابولا ویروسی 40 با استفاده از رزونانس پلاسما سطح
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
چکیده انگلیسی


- Single domain antibodies (sdAb) specific for VP40 from Ebola Virus were selected.
- Monoclonal and sdAb were evaluated using surface plasmon resonance.
- Monoclonal and sdAb had sub nM and nM dissociation constants respectively.
- An array format facilitated evaluation of the epitopes recognized by the antibodies.

Immunoassay formats, in which antibodies provide sensitivity and specificity, are often utilized to provide rapid and simple diagnostic tests. Surface plasmon resonance is frequently used to evaluate the suitability of antibodies by determining binding kinetics to agents or surrogate antigens. We used SPR to evaluate a number of commercial monoclonal antibodies as well as single domain antibodies produced in-house. All the antibodies targeted the Ebola virus viral protein 40 (VP40). We determined the ability of each antibody to bind to immobilized VP40, and ensured they did not bind Ebola glycoprotein or the nucleoprotein. A subset of the monoclonal antibodies was immobilized to characterize antigen capture in solution. It can be advantageous to utilize antibodies that recognize distinct epitopes when choosing reagents for detection and diagnostic assays. We determined the uniqueness of the epitope recognized by the anti-VP40 antibodies using a checkerboard format that exploits the 6 × 6 array of interactions monitored by the Bio-Rad ProteOn XPR36 SPR instrument. The results demonstrate the utility of surface plasmon resonance to characterize monoclonal and recombinant antibodies. Additionally, the analysis presented here enabled the identification of pairs of anti-VP40 antibodies which could potentially be utilized in sandwich type immunoassays for the detection of Ebola virus.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 442, March 2017, Pages 42-48
نویسندگان
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