title: Improving Trastuzumab's Stability Profile by Removing the Two Degradation Hotspots.
abstract: Stability of recombinant monoclonal antibodies (mAbs) is essential for their clinical application. The presence of the two degradation hotspots, namely, LC-Asn30 and HC-Asp102, in its complementary determinant regions prevents trastuzumab (Herceptin®) from being supplied in a drug product format of liquid formulation. To improve the stability, a new antibody was created by replacing the two residues with chemically similar amino acids of LC-Gln30 and HC-Glu102. This new mAb, named as T-mAb2, exhibited a simple and more uniform charge heterogeneity profile than T-mAb1, which is trastuzumab made in our laboratory, as displayed by the difference between their main peak area percentages (82.9% for T-mAb2 vs. 60.5% for T-mAb1). Computer modeling results, physicochemical and biological characterization, and stability profiling studies on T-mAb2 and T-mAb1 demonstrated that stability of T-mAb2 was significantly improved. In comparison with T-mAb1, although its in vitro human epidermal growth factor receptor 2 (HER2)-target binding activities were reduced slightly, in vivo tumor growth inhibiting activity was not affected, as demonstrated by the study results using the SKOV3 xenograft mouse model. Hence, a new anti-HER2 antibody was generated with improved stability that could be used to produce the drug product in liquid formulation for cost saving and more convenient usage.
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China; Beijing Mabworks Biotech Company Ltd., Economic-Technological Development Area, Beijing 101111, China.
authors.forename: Yuemei
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Beijing Mabworks Biotech Company Ltd., Economic-Technological Development Area, Beijing 101111, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Beijing Mabworks Biotech Company Ltd., Economic-Technological Development Area, Beijing 101111, China.
authors.forename: Shusheng
authors.initials: S
authors.lastname: Geng
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Chunmei
authors.initials: C
authors.lastname: Hou
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Xingyin
authors.initials: X
authors.lastname: Li
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Lang
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Chunxia
authors.initials: C
authors.lastname: Qiao
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China.
authors.forename: Jiannan
authors.initials: J
authors.lastname: Feng
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China. Electronic address: lm62033@163.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Lv
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, Beijing 100850, China. Electronic address: shenbf0714@163.com.
authors.forename: Beifen
authors.initials: B
authors.lastname: Shen
authors.affiliation: Beijing Mabworks Biotech Company Ltd., Economic-Technological Development Area, Beijing 101111, China. Electronic address: zhangby@mab-works.com.
authors.forename: Boyan
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000276:immunology / Q000627:therapeutic use; D001940:Breast / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D001943:Breast Neoplasms / Q000188:drug therapy / Q000276:immunology / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D055550:Protein Stability; D059748:Proteolysis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology; D000068878:Trastuzumab / Q000737:chemistry* / Q000276:immunology / Q000627:therapeutic use
chemical_list: D000970:Antineoplastic Agents; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human
keywords: HPLC; biotechnology; deamidation; mass spectrometry; molecular modeling; monoclonal antibody; physicochemical properties; stability; stabilization; trastuzumab