title: A novel method for detecting neutralizing antibodies against therapeutic proteins by measuring gene expression.
abstract: The presence of neutralizing antibodies against protein therapeutics is a concern in the biomedical field. Such antibodies not only reduce the efficacy of protein therapeutics, but also impose potential dangers to the patients receiving them. To date, a small number of in vitro cell-based bioassays for detecting neutralizing antibodies against therapeutic proteins have been developed. Most of the existing assays, however, either involve the use of radioactive materials or have limited sensitivities and/or poor specificities. With advances in mRNA profiling and detection techniques, we have established a novel and non-radioactive bioassay system using branched DNA (bDNA) technology for detecting protein-therapeutic neutralizing antibodies in patient serum. Our assay measures the variations of target gene expression that reflect the biologic effect of the therapeutic agent and the capability of the antibodies, if present, to neutralize the therapeutics. Compared with most existing assays, the new assay is more sensitive and specific, and completely eliminates the use of radioactive materials. Application of the new assay system can be widely expanded if new target genes and responding cell lines for other therapeutics are identified or engineered.
authors.affiliation: Department of Clinical Immunology, Amgen Inc, One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Yu
authors.forename: Christopher
authors.initials: C
authors.lastname: Piddington
authors.forename: Dan
authors.initials: D
authors.lastname: Fitzpatrick
authors.forename: Brian
authors.initials: B
authors.lastname: Twomey
authors.forename: Ren
authors.initials: R
authors.lastname: Xu
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Swanson
authors.forename: Shuqian
authors.initials: S
authors.lastname: Jing
mesh_terms: D000906:Antibodies / Q000097:blood* / Q000235:genetics; D000918:Antibody Specificity; D021121:Branched DNA Signal Amplification Assay / Q000379:methods*; D002070:Butadienes / Q000494:pharmacology; D045744:Cell Line, Tumor; D002867:Chromones / Q000494:pharmacology; D004921:Erythropoietin / Q000276:immunology / Q000494:pharmacology; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051573:Proto-Oncogene Proteins c-pim-1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D002070:Butadienes; D002867:Chromones; D009025:Morpholines; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; C113580:U 0126; D004921:Erythropoietin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D051573:Proto-Oncogene Proteins c-pim-1