The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains
Open accessAbstract
The display of proteins on the surface of phage offers a powerful means of selecting for rare genes encoding proteins with binding activities. Recently we found that antibody heavy and light chain variable (V) domains fused as a single polypeptide chain to a minor coat protein of filamentous phage fd, could be enriched by successive rounds of phage growth and panning with antigen. This allows the selection of antigen-binding domains directly from diverse libraries of V-genes. Now we show that heterodimeric Fab fragments can be assembled on the surface of the phage by linking one chain to the phage coat protein, and secreting the other into the bacterial periplasm. Furthermore by introducing an amber mutation between the antibody chain and the coat protein, we can either display the antibody on phage using supE strains of bacteria, or produce soluble Fab fragment using non-suppressor strains. The use of Fab fragments may offer advantages over single chain Fv fragments for construction of combinatorial libraries.
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Citations
Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961
Cited by
Montoyo, H. P., Vaccaro, C., Hafner, M., Ober, R. J., Mueller, W., & Ward, E. S. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0810796106 · Cited by 211
Comparative study of the p53-mdm2 and p53-MDMX interfaces
Böttger, V., Böttger, A., Garcia-Echeverria, C., Ramos, Y. F., van der Eb, A. J., Jochemsen, A. G., & Lane, D. P. (1999) · Oncogene · doi:10.1038/sj.onc.1202281 · Cited by 189
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
Tiede, C., Tang, A. A. S., Deacon, S. E., Mandal, U., Nettleship, J. E., Owen, R. L., George, S. E., Harrison, D. J., Owens, R. J., Tomlinson, D. C., et al. (2014) · Protein Engineering, Design and Selection · doi:10.1093/protein/gzu007 · Cited by 163
A Novel Heme Protein, the Cu,Zn-Superoxide Dismutase from Haemophilus ducreyi
Pacello, F., Langford, P. R., Kroll, J. S., Indiani, C., Smulevich, G., Desideri, A., Rotilio, G., & Battistoni, A. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m010488200 · Cited by 27
In Vitro Antibody Affinity Maturation Targeting Germline Hotspots
Ho, M., & Pastan, I. (2009) · Methods in Molecular Biology™ · doi:10.1007/978-1-59745-554-1_15 · Cited by 21
“Other comparable phagemids such as pHEN1 for scFv (12) and pComb3 for Fab (13) can be used as well.”
Method
Metadata
- Authors
- Hoogenboom, H. R., Griffiths, A. D., Johnson, K. S., Chiswell, D. J., Hudson, P., & Winter, G.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1991
- Volume
- 19
- Issue
- 15
- Pages
- 4133-4137
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2025827008
- Cited by
- 1,020
- References
- 1
- FWCI
- 41.20
- Type
- journal article
- Language
- en
- Open access
- bronze
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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