Abstract
A semisynthetic antibody library composed of single chain Fv fragments (scFv) was constructed by replacing the heavy chain CDR3 region of a human scFv by a random sequence of eight amino acids using trinucleotide codons. After cloning into a phage display vector, an antibody library was generated with a complexity of 8 x 10(8) independent clones. The library was screened for binders to dinitrophenol, fluorescein isothiocyanate and 3-nitro-4-hydroxy-5-iodophenylacetic acid. scFv antibodies that specifically bound the antigen were obtained in each case.
Full Text
The Full Text of this article is available as a PDF (75.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Akamatsu Y., Cole M. S., Tso J. Y., Tsurushita N. Construction of a human Ig combinatorial library from genomic V segments and synthetic CDR3 fragments. J Immunol. 1993 Nov 1;151(9):4651–4659. [PubMed] [Google Scholar]
- Clackson T., Hoogenboom H. R., Griffiths A. D., Winter G. Making antibody fragments using phage display libraries. Nature. 1991 Aug 15;352(6336):624–628. doi: 10.1038/352624a0. [DOI] [PubMed] [Google Scholar]
- Kayushin A. L., Korosteleva M. D., Miroshnikov A. I., Kosch W., Zubov D., Piel N. A convenient approach to the synthesis of trinucleotide phosphoramidites--synthons for the generation of oligonucleotide/peptide libraries. Nucleic Acids Res. 1996 Oct 1;24(19):3748–3755. doi: 10.1093/nar/24.19.3748. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Little M., Breitling F., Dübel S., Fuchs P., Braunagel M. Human antibody libraries in Escherichia coli. J Biotechnol. 1995 Jul 31;41(2-3):187–195. doi: 10.1016/0168-1656(95)00022-i. [DOI] [PubMed] [Google Scholar]
- Marks J. D., Griffiths A. D., Malmqvist M., Clackson T. P., Bye J. M., Winter G. By-passing immunization: building high affinity human antibodies by chain shuffling. Biotechnology (N Y) 1992 Jul;10(7):779–783. doi: 10.1038/nbt0792-779. [DOI] [PubMed] [Google Scholar]
- Marks J. D., Hoogenboom H. R., Bonnert T. P., McCafferty J., Griffiths A. D., Winter G. By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J Mol Biol. 1991 Dec 5;222(3):581–597. doi: 10.1016/0022-2836(91)90498-u. [DOI] [PubMed] [Google Scholar]
- Nissim A., Hoogenboom H. R., Tomlinson I. M., Flynn G., Midgley C., Lane D., Winter G. Antibody fragments from a 'single pot' phage display library as immunochemical reagents. EMBO J. 1994 Feb 1;13(3):692–698. doi: 10.1002/j.1460-2075.1994.tb06308.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welschof M., Terness P., Kipriyanov S. M., Stanescu D., Breitling F., Dörsam H., Dübel S., Little M., Opelz G. The antigen-binding domain of a human IgG-anti-F(ab')2 autoantibody. Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1902–1907. doi: 10.1073/pnas.94.5.1902. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Kruif J., Boel E., Logtenberg T. Selection and application of human single chain Fv antibody fragments from a semi-synthetic phage antibody display library with designed CDR3 regions. J Mol Biol. 1995 Apr 21;248(1):97–105. doi: 10.1006/jmbi.1995.0204. [DOI] [PubMed] [Google Scholar]