title: Small sized EGFR1 and HER2 specific bifunctional antibody for targeted cancer therapy.
abstract: Targeting tumors using miniature antibodies is a novel and attractive therapeutic approach, as these biomolecules exhibit low immunogenicity, rapid clearance, and high targeting specificity. However, most of the small-sized antibodies in existence do not exhibit marked anti-tumor effects, which limit their use in targeted cancer immunotherapy. To overcome this difficulty in targeting multiple biomarkers by combination therapies, we designed a new bifunctional antibody, named MaAbNA (multivalent antibody comprised of nanobody and affibody moieties), capable of targeting EGFR1 and HER2, which are widely overexpressed in a variety of tumor types. The small-sized (29 kDa) MaAbNA, which was expressed in E.coli, consists of one anti-EGFR1 nanobody and two anti-HER2 affibodies, and possesses high affinity (KD) for EGFR1 (~4.1 nM) and HER2 (~4.7 nM). In order to enhance its anti-tumor activity, MaAbNA was conjugated with adriamycin (ADM) using a PEG2000 linker, forming a new complex anticancer drug, MaAbNA-PEG2000-ADM. MaAbNA exhibited high inhibitory effects on tumor cells over-expressing both EGFR1 and HER2, but displayed minimal cytotoxicity in cells expressing low levels of EGFR1 and HER2. Moreover, MaAbNA-PEG2000-ADM displayed increased tumoricidal effects than ADM or MaAbNA alone, as well exhibited greater antitumor efficacy than EGFR1 (Cetuximab) and HER2 (Herceptin) antibody drugs. The ability of MaAbNA to regulate expression of downstream oncogenes c-jun, c-fos, c-myc, as well as AEG-1 for therapeutic potential was evaluated by qPCR and western-blot analyses. The antitumor efficacy of MaAbNA and its derivative MaAbNA-PEG2000-ADM were validated in vivo, highlighting the potential for use of MaAbNA as a highly tumor-specific dual molecular imaging probe and targeted cancer therapeutic.
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Ding
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Caiping
authors.initials: C
authors.lastname: Tian
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Song
authors.initials: S
authors.lastname: Feng
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Guissi
authors.initials: G
authors.lastname: Fida
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Congying
authors.initials: C
authors.lastname: Zhang
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Ma
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Guanhua
authors.initials: G
authors.lastname: Ai
authors.affiliation: 2. Department of Radiology, Washington University, School of Medicine, St. Louis, USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Achilefu
authors.affiliation: 1. Department of Biomedical Engineering, China Pharmaceutical University, School of Life Science and Technology, Nanjing, China.
authors.forename: Yueqing
authors.initials: Y
authors.lastname: Gu
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification* / Q000627:therapeutic use; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D009369:Neoplasms / Q000473:pathology / Q000628:therapy*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000627:therapeutic use; D061905:Single-Domain Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000627:therapeutic use; D016896:Treatment Outcome
chemical_list: D018033:Antibodies, Bispecific; D066246:ErbB Receptors; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; C512478:EGFR protein, human; C508053:ERBB2 protein, human
keywords: EGFR1; HER2