Anti-hepatoma human single-chain Fv antibody and adriamycin conjugates with potent antitumor activity.
To construct an improved biological missile, an immunoconjugate ADM-Dex-ScFv-SA3 was synthesized, which was composed of a hepatocellular carcinoma-specific, single-chain Fv antibody (ScFv-SA3) and a highly potent cytotoxic drug, adriamycin (ADM), as the warhead. Oxidized Dextran T10 (Dex-T10) was used as a linker to connect these two moieties. The 40 KD soluble anti-hepatoma human Trx-ScFv-SA3 protein was expressed in E. coli BL21 (DE3), using a prokaryotic expression vector, pET21a (+)-Trx-ScFv-SA3-His. It was purified using a His-Tag Ni-Agarose column and identified by western blot. The activity of Trx-ScFv-SA3 was verified by enzyme-linked immunosorbent assay (ELISA) and immunocytochemistry to confirm that it specifically binds to the hepatocellular carcinoma cell line HepG2. To prepare ADM-Dex-ScFv-SA3, ADM was conjugated to the antibody at a molar ratio of 14.21:1. The antitumor effect of the conjugate was tested by MTT assay, plate colony formation assay and xenografts in a nude mice experimental model. In vitro experiments revealed that ADM-Dex-ScFv-SA3 could bind to tumor cells selectively and inhibit the proliferation and the colony formation ability of HepG2 cells. In vivo experiments showed that ADM-Dex-ScFv-SA3 suppressed the tumor growth and prolonged the median survival time in tumor-bearing mice. Tumor histology slides indicated a significantly slower tumor tissue proliferation in the ADM-Dex-ScFv-SA3 group. These data indicate that the targeted drug, ADM-Dex-ScFv-SA3, may be a highly potent and selective therapy for the treatment of hepatoma.
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Lin
L
Chen
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Yan-Hong
YH
Liu
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Yue-Hui
YH
Li
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Yan
Y
Jiang
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Ping-Li
PL
Xie
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China.
Guo-Hua
GH
Zhou
Tumor Immunobiology Laboratory of Cancer Research Institute, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis and Cancer Invasion Ministry of Education, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China; Key Laboratory of Carcinogenesis Ministry of Health, Cancer Research Institution, Xiangya Medical School, Central South University, 410078 Changsha, Hunan, China. Electronic address: libsun@163.com.
Guan-Cheng
GC
Li
D000818:Animals; D000912:Antibodies, Neoplasm / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000628:therapy*; D049109:Cell Proliferation / Q000187:drug effects; D004317:Doxorubicin / Q000008:administration & dosage* / Q000737:chemistry; D005822:Genetic Vectors / Q000235:genetics; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D057127:Single-Chain Antibodies / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D047368:Tumor Burden / Q000187:drug effects; D014410:Tumor Stem Cell Assay; D023041:Xenograft Model Antitumor Assays
D000912:Antibodies, Neoplasm; D000970:Antineoplastic Agents; D057127:Single-Chain Antibodies; D004317:Doxorubicin
Adriamycin; Conjugate; Hepatocellular carcinoma; Single-chain Fv antibody