title: Site-specific DNA-antibody conjugates for specific and sensitive immuno-PCR.
abstract: Antibody conjugates are widely used as diagnostics and imaging reagents. However, many such conjugates suffer losses in sensitivity and specificity due to nonspecific labeling techniques. We have developed methodology to site-specifically conjugate oligonucleotides to antibodies containing a genetically encoded unnatural amino acid with orthogonal chemical reactivity. These oligobody molecules were used in immuno-PCR assays to detect Her2(+) cells with greater sensitivity and specificity than nonspecifically coupled fragments, and can detect extremely rare Her2(+) cells in a complex cellular environment. Such designed antibody-oligonucleotide conjugates should provide sensitive and specific reagents for diagnostics, as well as enable other unique applications based on oligobody building blocks.
authors.affiliation: Department of Chemistry, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Kazane
authors.forename: Devin
authors.initials: D
authors.lastname: Sok
authors.forename: Edward H
authors.initials: EH
authors.lastname: Cho
authors.forename: Maria Loressa
authors.initials: ML
authors.lastname: Uson
authors.forename: Peter
authors.initials: P
authors.lastname: Kuhn
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
authors.forename: Vaughn V
authors.initials: VV
authors.lastname: Smider
mesh_terms: D000906:Antibodies / Q000737:chemistry; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D006801:Humans; D007107:Immune System; D007700:Kinetics; D007962:Leukocytes / Q000166:cytology; D008856:Microscopy, Fluorescence / Q000379:methods; D009369:Neoplasms / Q000175:diagnosis; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D013696:Temperature
chemical_list: D000906:Antibodies; D004247:DNA; D009841:Oligonucleotides; D018719:Erb-b2 Receptor Tyrosine Kinases