دورية أكاديمية
Construction of a Fully Synthetic Human scFv Antibody Library with CDR3 Regions Randomized by a Split-Mix-Split Method and Its Application
| العنوان: | Construction of a Fully Synthetic Human scFv Antibody Library with CDR3 Regions Randomized by a Split-Mix-Split Method and Its Application |
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| المؤلفون: | Yin, Chang-Cheng, Ren, Li-Li, Zhu, Lin-Lin, Wang, Xiang-Bin, Zhang, Zhong, Huang, Hua-Liang, Yan, Xi-Yun |
| بيانات النشر: | Oxford University Press |
| سنة النشر: | 2008 |
| المجموعة: | HighWire Press (Stanford University) |
| مصطلحات موضوعية: | Regular Papers |
| الوصف: | The randomization scheme of hypervariable region takes crucial role in construction of a synthetic antibody library. The codon bias and inevitable ‘stop’ codon of conventional ‘NNK’ and ‘NNS’ codons limit their applications. Here we report a split–mix–split DNA synthesis method that can control over the amino acid composition and distribution of randomized sequences effectually. A fully synthetic human antibody library with a diversity of 1.56 × 109 was successfully generated with complementarity determining region 3 (CDR3) randomized by this strategy. Sequencing analysis indicated that >60% of colonies had completely correct scFv genes and the amino acid composition and distribution were designed well in accordance. The utility was demonstrated by screening of scFv clones against BHL (anti-CD3 × anti-ovarian carcinoma bispecific antibody). These results proved the feasibility of the split–mix–split DNA randomization strategy in library construction and site-directed mutagenesis. |
| نوع الوثيقة: | text |
| وصف الملف: | text/html |
| اللغة: | English |
| العلاقة: | http://jb.oxfordjournals.org/cgi/content/short/144/5/591Test; http://dx.doi.org/10.1093/jb/mvn103Test |
| DOI: | 10.1093/jb/mvn103 |
| الإتاحة: | https://doi.org/10.1093/jb/mvn103Test http://jb.oxfordjournals.org/cgi/content/short/144/5/591Test |
| حقوق: | Copyright (C) 2008, Japanese Biochemical Society |
| رقم الانضمام: | edsbas.AFAC9A93 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1093/jb/mvn103 |
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