title: Stability of the recombinant anti‑erbB2 scFv‑Fc‑interleukin‑2 fusion             protein and its inhibition of HER2‑overexpressing tumor cells.
abstract: The anti‑erbB2 scFv‑Fc‑IL‑2 fusion protein (HFI) is the basis for development of a novel targeted anticancer drug, in particular for the treatment of HER2‑positive cancer patients. HFI was fused with the anti‑erbB2 antibody and human IL‑2 by genetic engineering technology and by antibody targeting characteristics of HFI. IL‑2 was recruited to target cells to block HER2 signaling, inhibit or kill tumor cells, improve the immune capacity, reduce the dose of antibody and IL‑2 synergy. In order to analyse HFI drug ability, HFI plasmid stability was verified by HFI expression of the trend of volume changes. Additionally, HFI could easily precipitate and had progressive characteristics and thus, the buffer system of the additive phosphate‑citric acid buffer, arginine, Triton X‑100 or Tween‑80, the establishment of a microfiltration, ion exchange, affinity chromatography and gel filtration chromatography‑based purification process were explored. HFI samples were obtained according to the requirements of purity, activity and homogeneity. In vivo, HFI significantly delayed HER2 overexpression of non‑small cell lung cancer (Calu‑3) in human non‑small cell lung cancer xenografts in nude mice, and the inhibition rate was more than 60% (P<0.05) in the group treated with 1 mg/kg the HFI dose; HFI significantly inhibited HER2 expression of breast cancer (FVB/neu) transgenic mouse tumor growth in 1 mg/kg of the HFI dose group, and in the following treatment the 400 mm3 tumors disappeared completely. Combined with other HFI test data analysis, HFI not only has good prospects, but also laid the foundation for the development of antibody‑cytokine fusion protein‑like drugs.
authors.affiliation: Norman Bethune College of Medicine, Jilin University, Changchun                     130021, P.R. China.
authors.forename: Yu-Jia
authors.initials: YJ
authors.lastname: Du
authors.forename: Ze-Min
authors.initials: ZM
authors.lastname: Lin
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Zhao
authors.forename: Xiu-Ping
authors.initials: XP
authors.lastname: Feng
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Wang
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Chun-Di
authors.initials: CD
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Shi
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Zuo
authors.forename: Fan
authors.initials: F
authors.lastname: Li
authors.forename: Cheng-Zhong
authors.initials: CZ
authors.lastname: Wang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000473:pathology; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007376:Interleukin-2 / Q000235:genetics* / Q000276:immunology; D061986:MCF-7 Cells; D051379:Mice; D055550:Protein Stability; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics*; D015398:Signal Transduction; D023041:Xenograft Model Antitumor Assays
chemical_list: D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D007135:Immunoglobulin Variable Region; D007376:Interleukin-2; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; C508594:IL2 protein, human; C508053:ERBB2 protein, human