A fast and efficient procedure to produce scFvs specific for large macromolecular complexes.
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.
Wu
Si
S
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
Ke
Ailong
A
Doudna
Jennifer A
JA
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
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