title: Biosensing Probe for Quality Control Monitoring of the Structural Integrity of Therapeutic Antibodies.
abstract: Ideal quality control of therapeutic antibodies involves analytical techniques with high-sensitivity, high-resolution, and high-throughput performance. Few technologies that assess the physicochemical heterogeneity of antibodies, however, meet all the required demands. We developed a biosensing method for the quality control of therapeutic antibodies based on an artificial protein, AF.2A1, which discriminates between the native and the non-native three-dimensional structures of immunoglobulin G (IgG). AF.2A1 specifically recognized non-native IgG spiked into a solution of native IgG, thereby making it possible to detect contamination by a small amount of non-native IgG, which is difficult using conventional size-based separation or spectroscopic techniques. Using AF.2A1 as an analytical probe, we determined the concentration of non-native IgG formed under various pH conditions. The probe was also applicable to accelerated tests of the long-term stability of a therapeutic antibody, allowing monitoring of the formation of non-native IgG at elevated temperatures and extended periods of storage. AF.2A1, a proteinous probe, can be combined with established methods or devices to achieve high-throughput assays and provides the potential for probe-based biosensing for the quality control of therapeutic antibodies.
authors.affiliation: Biomedical Research Institute, the National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Watanabe
authors.affiliation: Biomedical Research Institute, the National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.|Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, the University of Tokyo , 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan.
authors.forename: Seiki
authors.initials: S
authors.lastname: Yageta
authors.affiliation: Biomedical Research Institute, the National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Imamura
authors.affiliation: Biomedical Research Institute, the National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.|Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, the University of Tokyo , 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Honda
mesh_terms: D000906:Antibodies / Q000032:analysis*; D061067:Antibodies, Monoclonal, Humanized / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods; D007074:Immunoglobulin G / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry*; D011786:Quality Control
chemical_list: D000906:Antibodies; D061067:Antibodies, Monoclonal, Humanized; D007074:Immunoglobulin G; D011506:Proteins