Phage display of single-chain antibody constructs.
Phage display is based on expressing recombinant proteins fused to a phage coat protein. The genetic information encoding for the displayed molecule is physically linked to its product via the displaying particle. With phage display, tailor-made proteins (fused peptides, antibodies, enzymes, DNA-binding proteins) may be synthesized and selected to acquire the desired catalytic properties or affinity and specificity of binding. This unit provides protocols for the construction of, and selection from antibody phage display libraries, together with useful approaches for the analysis of phage antibodies. This unit focuses on the construction of immune libraries where the sources for antibody genes are spleens of immunized mice and the isolation and initial characterization of single-chain antibodies (scFvs) from such libraries.
Benhar
Itai
I
The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Ramat Aviv, Israel.
Reiter
Yoram
Y
D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics*; D001710:Biotin / Q000031:analogs & derivatives / Q000378:metabolism; D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000378:metabolism; D016172:DNA Fingerprinting; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000821:virology; D029968:Escherichia coli Proteins / Q000378:metabolism; D005822:Genetic Vectors; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D051379:Mice; D010969:Plastics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D012097:Repressor Proteins / Q000378:metabolism; D013388:Succinimides / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D000906:Antibodies; D000941:Antigens; D029968:Escherichia coli Proteins; D007135:Immunoglobulin Variable Region; D010969:Plastics; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D013388:Succinimides; D014157:Transcription Factors; C053827:biotinyl-N-hydroxysulfosuccinimide ester; D001710:Biotin; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli