A recombinant, fully human monoclonal antibody with antitumor activity constructed from phage-displayed antibody fragments

Huls, Gerwin; Heijnen, Ingmar; Cuomo, Maria Emanuela; Koningsberger, J. C.; Wiegman, Luus; Boel, Edwin; Vries, Anne-Renée Van der Vuurst de; Loyson, Sabine A.J.; Helfrich, Wijnand; Henegouwen, G. P. van Berge; Meijer, Marja van; Kruif, John de; Logtenberg, Ton

doi:10.1038/7023

Cited by 93 publications

(55 citation statements)

References 40 publications

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“…48 Interestingly, some reports have indicated that HAMA has little adverse effect on the overall clinical outcome of the mAb therapy; 49,50 however, reduction in toxicity will be of benefit as would increased persistence of the immune conjugate. Our use of a human IgG should limit immune reaction against the fusion protein 51 and although the scFv is murine, techniques to humanize this part of the construct such as the use of human phage libraries 52 could be used. In addition, genetic delivery of the immune conjugates will take full advantage of the relatively small size of the molecules for good tumor penetration and should also be able to circumvent the potential disadvantages of rapid clearance from the blood plasma.…”

Section: Discussionmentioning

confidence: 99%

Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins

Myers

Ryan

Stern³

et al. 2002

Cancer Gene Ther

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Although several clinical trials have shown beneficial effects by targeting tumor -associated antigens ( TAAs ) with monoclonal antibodies, a number of issues, including poor penetration of the tumor mass and human antimouse antibody responses, remain. The use of recombinant single -chain Fv ( scFv ) fragments has the potential to address these and other issues while allowing the addition of different effector functions. To develop therapeutic strategies that recruit both humoral and cellular arms of the immune response, we have constructed chimeric proteins linking either the human IgG1 Fc domain or the extracellular domain of murine B7.1 to a scFv specific for the oncofetal glycoprotein, 5T4. This TAA is expressed by a wide variety of carcinomas and is associated with metastasis and poorer clinical outcome. We have engineered retroviral constructs that produce fusion proteins able to interact simultaneously with both 5T4 -positive cells and with the receptor / ligands of the immune effector moieties. Genetic delivery through a murine leukemia virus vector to 5T4 -positive tumor cells results in the secreted scFv fusion protein binding to the cell surface. Furthermore, the scFv -HIgG1 fusion protein is able to direct lysis of 5T4 -expressing human tumor cell lines through antibody -dependent cell cytotoxicity, indicating its potential as a gene therapy for human cancers.

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Section: Discussionmentioning

confidence: 99%

Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins

Myers

Ryan

Stern³

et al. 2002

Cancer Gene Ther

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“…The anti-Ep-CAM scFv C28 was derived from scFv UBS-54, which was isolated from a semi-synthetic phage antibody display library in a pHEN vector (Huls et al, 1999). An SfiI/NotI fragment encoding the scFv C28 was isolated from the pHEN vector and cloned into the eukaryotic expression vector, pSTCF (Arafat et al, 2000), containing a secretion signal and a myc-and 6His-tag.…”

Section: Construction Of Pc28-gushmentioning

confidence: 99%

A fully human anti-Ep-CAM scFv-beta-glucuronidase fusion protein for selective chemotherapy with a glucuronide prodrug

et al. 2002

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Monoclonal antibodies against tumour-associated antigens could be useful to deliver enzymes selectively to the site of a tumour for activation of a non-toxic prodrug. A completely human fusion protein may be advantageous for repeated administration, as host immune responses may be avoided. We have constructed a fusion protein consisting of a human single chain Fv antibody, C28, against the epithelial cell adhesion molecule and the human enzyme b-glucuronidase. The sequences encoding C28 and human enzyme b-glucuronidase were joined by a sequence encoding a flexible linker, and were preceded by the IgGk signal sequence for secretion of the fusion protein. A CHO cell line was engineered to secrete C28-bglucuronidase fusion protein. Antibody specificity and enzyme activity were retained in the secreted fusion protein that had an apparent molecular mass of 100 kDa under denaturing conditions. The fusion protein was able to convert a non-toxic prodrug of doxorubicin, N-

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“…Having taken this into consideration (33,38), the in vitro cytotoxicity of anti-EpCAM(scFv)-MAP was determined using an XTT cell viability assay. Anti-EpCAM(scFv)-MAP showed specific toxicity toward L3.6pl and A431 cells reaching IC 50 values of 43 nmol/L and 67 nmol/L, respectively (Fig.…”

Section: Cytotoxicity Of Anti-epcam(scfv)-map In Vitromentioning

confidence: 99%

EpCAM-Selective Elimination of Carcinoma Cells by a Novel MAP-Based Cytolytic Fusion Protein

Hristodorov

Amoury

Mladenov

et al. 2014

Molecular Cancer Therapeutics

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In normal epithelia, the epithelial cell adhesion molecule (EpCAM) expression is relatively low and only present at the basolateral cell surface. In contrast, EpCAM is aberrantly overexpressed in various human carcinomas. Therefore, EpCAM is considered to be a highly promising target for antibody-based cancer immunotherapy. Here, we present a new and fully human cytolytic fusion protein (CFP), designated "antiEpCAM(scFv)-MAP," that is comprised of an EpCAM-specific antibody fragment (scFv) genetically fused to the microtubule-associated protein tau (MAP). Anti-EpCAM(scFv)-MAP shows potent EpCAM-restricted proapoptotic activity toward rapidly proliferating carcinoma cells. In vitro assays confirmed that treatment with anti-EpCAM(scFv)-MAP resulted in the colocalization and stabilization of microtubules, suggesting that this could be the potential mode of action. Dose-finding experiments indicated that anti-EpCAM(scFv)-MAP is well tolerated in mice. Using noninvasive far-red in vivo imaging in a tumor xenograft mouse model, we further demonstrated that anti-EpCAM(scFv)-MAP inhibited tumor growth in vivo. In conclusion, our data suggest that anti-EpCAM(scFv)-MAP may be of therapeutic value for the targeted elimination of EpCAM þ carcinomas. Mol Cancer Ther; 13(9); 2194-202. Ó2014 AACR.

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A recombinant, fully human monoclonal antibody with antitumor activity constructed from phage-displayed antibody fragments

Cited by 93 publications

References 40 publications

Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins

Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins

A fully human anti-Ep-CAM scFv-beta-glucuronidase fusion protein for selective chemotherapy with a glucuronide prodrug

EpCAM-Selective Elimination of Carcinoma Cells by a Novel MAP-Based Cytolytic Fusion Protein

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