The intracellular antibody capture technology (IACT): towards a consensus sequence for intracellular antibodies.
We describe the application of an intracellular antibody capture technology (IACT) as a generic in vivo selection procedure for isolating intracellular antibodies or ICAbs. IACT was applied to the de novo selection of functional ICAbs against the microtubule-associated protein TAU, found in neurofibrillary lesions of Alzheimer's disease brains. A panel of 17 different ICAbs was created which bind TAU inside cells and the epitopes recognized by the selected ICAbs have been determined by an in vivo epitope mapping procedure. Finally, sequence analysis showed that the IACT-derived ICAbs are characterized by a common signature of conserved amino acid residues, suggesting that the IACT naturally selects a sort of "captured consensus sequence" for intracellular antibodies. The development of IACT, together with the possibility of scaling up in a high throughput and automated format, makes IACT a new enabling tool for target validation in functional genomics and global proteomics.
Visintin
Michela
M
International School for Advanced Studies (SISSA) and INFM Unit, 34013 Trieste, Italy.
Settanni
Giovanni
G
Maritan
Amos
A
Graziosi
Sergio
S
Marks
James D
JD
Cattaneo
Antonino
A
D000544:Alzheimer Disease / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity / Q000235:genetics* / Q000276:immunology*; D001331:Automation; D016466:CHO Cells; D019556:COS Cells; D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods*; D016384:Consensus Sequence / Q000235:genetics; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D006801:Humans; D007424:Intracellular Fluid / Q000276:immunology*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009154:Mutation; D019151:Peptide Library; D015203:Reproducibility of Results; D016415:Sequence Alignment; D016875:tau Proteins / Q000235:genetics / Q000276:immunology
D000906:Antibodies; D019151:Peptide Library; D016875:tau Proteins