| Recombinant single-chain variable fragment antibodies against extracellular epitopes of human multidrug resistance protein MRP3 for targeting malignant gliomas. | |
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MedLine Citation:
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PMID: 19937796 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Multidrug resistance protein 3 (MRP3), a multidrug resistance protein identified by serial analysis of gene expression as a glioblastoma multiforme (GBM)-associated molecule, is highly expressed in GBM, but not in normal brain cells. Thus, MRP3 is a candidate for GBM immunotargeting, but to date, no monoclonal antibody has been isolated that can target an extracellular MRP3 epitope. By phage display, we have isolated 3 recombinant, fully human, single-chain Fv (scFv) antibodies, M25, M58 and M89, which specifically react with the extracellular N-terminus of human MRP3. In ELISA, these scFvs reacted only with the peptide used for screening and not with other MRP3-derived peptides. Flow cytometric analysis revealed that these scFv fragments bind specifically to viable human GBM cells displaying different MRP3 expression levels, but not to MRP3-null cells. Furthermore, these scFv antibodies failed to react with tumor cells overexpressing other MRP proteins, including MRP1, MRP2, MRP4 and MRP5. M25 and M58 also bound to viable neurospheres. Iodogen-labeled scFvs demonstrated a yield of 56-76%. The immunoreactive fractions of the radiolabeled M25, M58 and M89 scFvs were 32, 52 and 69%, respectively. M25 exhibited 20% internalization into D2159MG neurospheres, M58, 33% into D54MG cells and M89, 26% into D247MG. Immunohistochemical evaluation of human gliomas to determine the localization of MRP3 antigen using scFvs M25 and M58 showed a dense cytoplasmic and membranous staining pattern. These Fv-based recombinant antibodies, which possess superior tumor penetration capabilities and selectively target tumor cells that express MRP3, may potentially be used in immunotherapy and diagnosis for brain tumors and other cancers. |
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Authors:
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Chien-Tsun Kuan; Nidhi Srivastava; Roger E McLendon; Wayne A Marasco; Michael R Zalutsky; Darell D Bigner |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: International journal of cancer. Journal international du cancer Volume: 127 ISSN: 1097-0215 ISO Abbreviation: Int. J. Cancer Publication Date: 2010 Aug |
Date Detail:
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Created Date: 2010-05-31 Completed Date: 2010-07-05 Revised Date: 2012-02-03 |
Medline Journal Info:
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Nlm Unique ID: 0042124 Medline TA: Int J Cancer Country: United States |
Other Details:
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Languages: eng Pagination: 598-611 Citation Subset: IM |
Affiliation:
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Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA. kuan@duke.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Antibody Specificity Base Sequence Brain Neoplasms / immunology*, pathology Cell Line, Tumor DNA Primers Enzyme-Linked Immunosorbent Assay Epitopes / immunology* Flow Cytometry Glioblastoma / immunology*, pathology Humans Immunoglobulin Fragments / immunology* Immunohistochemistry Molecular Sequence Data Multidrug Resistance-Associated Proteins / chemistry, immunology* |
| Grant Support | |
ID/Acronym/Agency:
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5P50 CA108786/CA/NCI NIH HHS; 5P50 NS20023/NS/NINDS NIH HHS; CA 74886/CA/NCI NIH HHS; MO1 RR30/RR/NCRR NIH HHS; P50 CA108786-05/CA/NCI NIH HHS; P50 NS020023-26/NS/NINDS NIH HHS; R37 CA 011898/CA/NCI NIH HHS; R37 CA011898-39/CA/NCI NIH HHS; R37 CA011898-41/CA/NCI NIH HHS; R37 CA011898-42/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/Epitopes; 0/Immunoglobulin Fragments; 0/Multidrug Resistance-Associated Proteins; 0/multidrug resistance-associated protein 3 |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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