title: Immobilization of a human epidermal growth factor receptor 2 mimotope-derived synthetic peptide on Au and its potential application for detection of herceptin in human serum by quartz crystal microbalance.
abstract: Therapeutic antibodies are antigenically similar to human antibodies and are difficult to detect in assays of human serum samples without the use of the therapeutic antibody's complementary antigen. Herein for the first time, we established a platform to detect Herceptin in solutions by using a small (<2.2 kDa), inexpensive, highly stable human epidermal growth factor receptor (HER2) mimotope-derived synthetic peptide immobilized on the surface of a Au quartz electrode. We used the HER2 mimotope as a substitute for the HER2 receptor protein in piezoimmunosensor or quartz crystal microbalance (QCM) assays to detect Herceptin in human serum. We demonstrated that assay sensitivity was dependent upon the amino acids used to tether and link the peptide to the sensor surface and the buffers used to carry out the assays. The detection limit of the piezoimmunosensor assay was 0.038 nM with a linear operating range of 0.038-0.859 nM. Little nonspecific binding to other therapeutic antibodies (Avastin and Rituxan) was observed. Levels of Herceptin in serum samples obtained from treated patients, as ascertained using the synthetic peptide-based QCM assay, were typical for those treated with Herceptin. The findings of this study are significant in that low-cost synthetic peptides could be used in a QCM assay, in lieu of native or recombinant antigens or capture antibodies, to rapidly detect a therapeutic antibody in human serum. The results suggested that a synthetic peptide bearing a particular functional sequence could be applied for developing a new generation of affinity-based immunosensors to detect a broad range of clinical biomarkers.
authors.affiliation: Chemistry Department, Oakland University, Rochester, Michigan 48309, USA.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Shang
authors.forename: Pankaj R
authors.initials: PR
authors.lastname: Singh
authors.forename: Mohammad M
authors.initials: MM
authors.lastname: Chisti
authors.forename: Ray
authors.initials: R
authors.lastname: Mernaugh
authors.forename: Xiangqun
authors.initials: X
authors.lastname: Zeng
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000097:blood*; D004566:Electrodes; D005260:Female; D006046:Gold / Q000737:chemistry*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D007118:Immunoassay; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D058525:Quartz Crystal Microbalance Techniques; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D006046:Gold; D055767:Immobilized Proteins; D010455:Peptides; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human