title: A fast and efficient procedure to produce scFvs specific for large macromolecular complexes.
abstract: We have expanded the application of antibody phage display to a new type of antigen: ribonucleoprotein (RNP) complexes. We describe a simple and efficient method for screening antibodies specific for large intact RNPs and individual components. We also describe a fast and easy method to overcome the abundance of amber stop codons in the positive phage clones. The resulting antibodies have been used in ELISA and Western blot analysis.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
authors.forename: Si
authors.initials: S
authors.lastname: Wu
authors.forename: Ailong
authors.initials: A
authors.lastname: Ke
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doudna
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology; D015153:Blotting, Western; D003090:Coliphages / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins / Q000276:immunology; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D046911:Macromolecular Substances / Q000737:chemistry / Q000276:immunology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000276:immunology; D018000:Receptors, Peptide / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D017422:Sequence Analysis, DNA; D018271:Signal Recognition Particle / Q000737:chemistry / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C082931:Ffh protein, E coli; C087406:FtsY protein, Bacteria; D007135:Immunoglobulin Variable Region; D046911:Macromolecular Substances; D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide; D011994:Recombinant Proteins; D018271:Signal Recognition Particle; C036318:signal peptide receptor