کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9902330 | 1545800 | 2005 | 16 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Construction, evaluation and refinement of a large human antibody phage library based on the IgD and IgM variable gene repertoire
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کلمات کلیدی
GFP2-phenyloxazol-5-onescFvMDAOMVPBLGPiVZV3,4-Methylenedioxyamphetamine - 3،4-MethylenedioxyamphetamineBSA - BSASingle chain Fv - Fv تنها زنجیره ایbovine serum albumin - آلبومین سرم گاوOuter membrane vesicle - غشای بیرونی vesiclePhox - فاکسperipheral blood lymphocytes - لنفوسیت های خون محیطیVariable - متغیرPhage display - نمایش فاژیVaricella zoster virus - ویروس واریسلا زوسترProtein L - پروتئین Lgreen fluorescent protein - پروتئین فلورسنت سبزAntibody library - کتابخانه آنتیبادیGlucose-6-phosphate isomerase - گلوکز 6-فسفات ایزومراز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The ability to isolate antibodies against any antigen of interest has become increasingly important as antibodies have proved their utility both in antigen detection, quantification and as specific in vivo targeting agents. To this end, we have constructed a large antibody phage library in the single chain Fv (scFv) phagemid format based on the naïve human variable (V) gene repertoire dictated by IgD and IgM. Optimizing each step of the library construction has resulted in a highly diverse and functional library, as assessed by sequencing analysis, large-scale automated expression analysis and antigen screening. Furthermore, the versatile format of the library, which comprises 14 separate sub-libraries, adds considerably flexibility with respect to which part of the antibody repertoire that is to be probed. This versatility has been further exploited to generate a refined antibody library, which exhibits one of the highest prokaryotic expression levels reported to date for a naïve repertoire. The construction of the refined library was based on the functional purification of expressed V genes in the context of the protein L interaction with correctly folded V genes of the kappa light chain family. Antigen screening of this library indicated that the functional purification improved the ability to retrieve antigen specific antibodies, but at the cost of potential loss of diversity in the isolated repertoire.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 299, Issues 1â2, April 2005, Pages 47-62
Journal: Journal of Immunological Methods - Volume 299, Issues 1â2, April 2005, Pages 47-62
نویسندگان
Geir Ã
ge Løset, Inger Løbersli, Anita Kavlie, John Eric Stacy, Tine Borgen, Louise Kausmally, Else Hvattum, Bjørg Simonsen, Maria Befring Hovda, Ole Henrik Brekke,