Generation of Antibody Libraries for Phage Display: Preparation of Helper Phage.
| Autor: | Peng H; Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, Florida 33458, USA., Rader C; Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, Florida 33458, USA rader33458@gmail.com. |
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| Jazyk: | angličtina |
| Zdroj: | Cold Spring Harbor protocols [Cold Spring Harb Protoc] 2024 Oct 15. Date of Electronic Publication: 2024 Oct 15. |
| DOI: | 10.1101/pdb.prot108600 |
| Abstrakt: | The generation and selection of antibody libraries by phagemid-based phage display requires three components; namely, phagemid library, host bacterial cells, and helper phage. The use of helper phage is necessary for the selection of phagemid libraries by phage display because it provides all genes needed for production of infectious phage particles. Here, we describe the generation of high-titer helper phage preparations suitable for phagemid-based phage display. The approach is based on helper phage VCSM13, which includes a gene for kanamycin resistance and a mutated packaging signal that, in the presence of a phagemid with an unmutated packaging signal, favors the production of infectious phage particles with phagemid phenotype and genotype. (© 2024 Cold Spring Harbor Laboratory Press.) |
| Databáze: | MEDLINE |
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