Quality of Recombinant Protein Determines the Amount of Autoreactivity Detected against the Tumor-Associated Epithelial Cell Adhesion Molecule Antigen: Low Frequency of Antibodies against the Natural Protein

Schmetzer, Oliver; Moldenhauer, Gerhard; Riesenberg, Rainer; Pires, Juliana Rico; Schlag, Peter M.; Pezzutto, Antonio

doi:10.4049/jimmunol.174.2.942

Cited by 21 publications

(22 citation statements)

References 43 publications

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“…ChCE7-F(abV) fragments were produced as histidine-tagged proteins in HEK293 cells and purified as described (24). HEA125, a mouse IgG1 directed against EpCAM, was described before and binds to all human adenocarcinomas (25). Anti-His antibody (tetra-His antibody) was purchased from Qiagen AG (Hombrechtikon, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

Efficient Inhibition of Intra-Peritoneal Tumor Growth and Dissemination of Human Ovarian Carcinoma Cells in Nude Mice by Anti-L1-Cell Adhesion Molecule Monoclonal Antibody Treatment

et al. 2006

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The L1 cell adhesion molecule is implicated in the control of proliferation, migration, and invasion of several tumor cell types in vitro. Recently, L1 overexpression was found to correlate with tumor progression of ovarian carcinoma, one of the most common causes of cancer-related deaths in gynecologic malignant diseases. To evaluate L1 as a potential target for ovarian cancer therapy, we investigated the effects of anti-L1 monoclonal antibodies (chCE7 and L1-11A) on proliferation and migration of L1-positive human SKOV3ip ovarian carcinoma cells in vitro and the therapeutic efficacy of L1-11A against i.p. SKOV3ip tumor growth in nude mice. In vitro, both anti-L1 antibodies efficiently inhibited the proliferation of SKOV3ip cells as well as other L1-expressing tumor cell lines (renal carcinoma, neuroblastoma, and colon carcinoma). On two cell lines, hyper-cross-linking of L1-11A with a secondary antibody was necessary for significant inhibition of proliferation, indicating that cross-linking of L1 is required for the antiproliferative effect. L1-negative prostate carcinoma cells were not influenced by antibody treatment. Biweekly treatment of ovarian carcinoma-bearing mice with L1-11A led to a dose-dependent and significant reduction of tumor burden (up to À63.5%) and ascites formation (up to À75%). This effect was associated with reduced proliferation within the tumors. L1-directed antibody-based inhibition of peritoneal growth and dissemination of human ovarian carcinoma cells represents important proof-of-principle for the development of a new therapy against one of the leading gynecologic malignant diseases. (Cancer Res 2006; 66(2): 936-43)

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Section: Methodsmentioning

confidence: 99%

Efficient Inhibition of Intra-Peritoneal Tumor Growth and Dissemination of Human Ovarian Carcinoma Cells in Nude Mice by Anti-L1-Cell Adhesion Molecule Monoclonal Antibody Treatment

et al. 2006

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“…Correlation of patient sample results by capture ELISA and indirect ELISA is significant antigen avoids many of the issues associated with developing high throughput assays for humoral immune monitoring such as lack of commercially prepared proteins and background responses to E. coli used in producing recombinant proteins [7,8]. We have previously shown that tumor cell lysate-based assays can be made CLIAready and suitable for high-throughput use in the clinical lab [6].…”

Section: Discussionmentioning

confidence: 99%

His-tag ELISA for the Detection of Humoral Tumor-Specific Immunity

Goodell

McNeel

Nguyen

et al. 2004

Journal of Immunotherapy

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Background: The application of high throughput molecular techniques such as SEREX are resulting in the identification of a multitude of tumor associated antigens. As newly identified antigens are incorporated into a variety of clinical trials, standardization of immunologic monitoring methods becomes increasingly important. We questioned whether mammalian cell expression of a histadine-linked human protein could be used to produce antigen suitable for detecting tumorspecific humoral immunity and whether such an assay could be amenable to standardization for clinical use.

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“…Sera from mice immunized with recombinant GA733-2 from baculovirusinfected insect cell systems show a binding activity to SW1116 colorectal carcinoma cells [4]. Recombinant GA733-2 proteins produced in Drosophila systems have also been used for detection of auto-antibodies [5]; however, the in vivo immunogenicity of recombinant GA733-2 derived from Drosophila systems in mice has not been reported.…”

Section: Introductionmentioning

confidence: 99%

“…Expression of recombinant GA733-2 has been reported in expression systems including baculovirus-infected Sf21 cells [4], Drosophila melanogaster S2 cells [5], and plant cell systems [6,7]. Sera from mice immunized with recombinant GA733-2 from baculovirusinfected insect cell systems show a binding activity to SW1116 colorectal carcinoma cells [4].…”

Section: Introductionmentioning

confidence: 99%

Expression and Immunogenicity of a Recombinant Chimeric Protein of Human Colorectal Cancer Antigen GA733-2 and an Fc Antibody Fragment in Stably Transformed Drosophila melanogaster S2 Cells

Seok

Kim

Yoo

et al. 2010

Appl Biochem Biotechnol

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Human colorectal cancer antigen GA733-2 fused to the immunoglobin Fc fragment (GA733-2-Fc) was expressed in stably transformed Drosophila melanogaster S2 cells, and the immunogenicity of recombinant GA733-2-Fc was investigated in mice. Recombinant GA733-2-Fc was secreted into a culture medium with a molecular mass of approximately 65 kDa. Recombinant GA733-2-Fc was purified to homogeneity using affinity fractionation with Protein A sepharose 4 Fast Flow. Recombinant GA733-2-Fc proteins elicited production of specific antibodies against recombinant GA733-2 by immunization through an intraperitoneal route. Recombinant GA733-2-Fc-induced antibodies showed a binding activity to human colorectal carcinoma HCT-116 cells. Secretory recombinant GA733-2-Fc from Drosophila S2 cell systems can be used as an effective experimental antigen for research in cancer vaccine development.

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Quality of Recombinant Protein Determines the Amount of Autoreactivity Detected against the Tumor-Associated Epithelial Cell Adhesion Molecule Antigen: Low Frequency of Antibodies against the Natural Protein

Cited by 21 publications

References 43 publications

Efficient Inhibition of Intra-Peritoneal Tumor Growth and Dissemination of Human Ovarian Carcinoma Cells in Nude Mice by Anti-L1-Cell Adhesion Molecule Monoclonal Antibody Treatment

Efficient Inhibition of Intra-Peritoneal Tumor Growth and Dissemination of Human Ovarian Carcinoma Cells in Nude Mice by Anti-L1-Cell Adhesion Molecule Monoclonal Antibody Treatment

His-tag ELISA for the Detection of Humoral Tumor-Specific Immunity

Expression and Immunogenicity of a Recombinant Chimeric Protein of Human Colorectal Cancer Antigen GA733-2 and an Fc Antibody Fragment in Stably Transformed Drosophila melanogaster S2 Cells

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