![](/images/graphics-bg.png)
In Vitro Selection of Fab Fragments by mRNA Display and Gene-Linking Emulsion PCR
Joint Authors
Yanagawa, Hiroshi
Doi, Nobuhide
Sumida, Takeshi
Source
Issue
Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2012-09-23
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
In vitro selection by display methods has been an effective tool for engineering recombinant antibodies.
mRNA display based on a cell-free translation system has the advantages of larger library sizes and quicker selection procedures compared with cell-based display methods such as phage display.
However, mRNA display has been limited to select single-chain polypeptides such as scFvs due to its characteristic of linking a nascent polypeptide with its encoding mRNA on the ribosome.
Here we demonstrated a new way of selecting heterodimeric Fab fragments by using mRNA display combined with emulsion PCR.
We designed a pair of complementary 5′ UTR sequences that can link the Fab heavy and light chain genes together by overlap-extension PCR in water-in-oil emulsions.
We confirmed that two mRNA-displayed polypeptides for heavy and light chain of a model Fab fragment were associated into the active form and that a specific Fab fragment gene was enriched over 100-fold per round of a model affinity selection followed by the gene-linking emulsion PCR.
We further performed directed evolution of Fab fragments with higher binding activity from a randomized Fab fragment library.
American Psychological Association (APA)
Sumida, Takeshi& Yanagawa, Hiroshi& Doi, Nobuhide. 2012. In Vitro Selection of Fab Fragments by mRNA Display and Gene-Linking Emulsion PCR. Journal of Nucleic Acids،Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-466748
Modern Language Association (MLA)
Sumida, Takeshi…[et al.]. In Vitro Selection of Fab Fragments by mRNA Display and Gene-Linking Emulsion PCR. Journal of Nucleic Acids No. 2012 (2012), pp.1-9.
https://search.emarefa.net/detail/BIM-466748
American Medical Association (AMA)
Sumida, Takeshi& Yanagawa, Hiroshi& Doi, Nobuhide. In Vitro Selection of Fab Fragments by mRNA Display and Gene-Linking Emulsion PCR. Journal of Nucleic Acids. 2012. Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-466748
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-466748