Simplified synthetic antibody libraries.
Synthetic antibody libraries are constructed from scratch using designed synthetic DNA. Precise control over design enables the use of highly optimized human frameworks and the introduction of defined chemical diversity at positions that are most likely to contribute to antigen recognition. We describe complete methods for the design, construction, and application of simplified synthetic antibody libraries built on a single human framework with diversity restricted to four complementarity-determining regions and two amino acids (tyrosine and serine). Despite the extreme simplicity of design, these libraries are capable of generating specific antibodies against diverse protein antigens. Moreover, the same methods can be used to build more complex libraries that can produce synthetic antibodies with affinities and specificities beyond the scope of natural antibodies. Most importantly, these simplified methods rely on standard supplies, equipment, and methods that are accessible to any molecular biology laboratory.
Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
Saravanan
S
Rajan
Sachdev S
SS
Sidhu
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics / Q000276:immunology*; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000915:Antibody Affinity / Q000276:immunology; D000941:Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics*; D019921:Biotinylation; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial
D000596:Amino Acids; D000906:Antibodies; D000941:Antigens; D019151:Peptide Library