Detection of amino-terminal extracellular domain of somatostatin receptor 2 by specific monoclonal antibodies and quantification of receptor density in medulloblastoma.
| dc.contributor.author | Kuan, Chien-Tsun | |
| dc.contributor.author | Wikstrand, Carol J | |
| dc.contributor.author | McLendon, Roger E | |
| dc.contributor.author | Zalutsky, Michael R | |
| dc.contributor.author | Kumar, Ujendra | |
| dc.contributor.author | Bigner, Darell D | |
| dc.coverage.spatial | United States | |
| dc.date.accessioned | 2011-04-15T16:46:29Z | |
| dc.date.issued | 2009-12 | |
| dc.description.abstract | Somatostatin receptor 2 (SSTR2) is expressed by most medulloblastomas (MEDs). We isolated monoclonal antibodies (MAbs) to the 12-mer (33)QTEPYYDLTSNA(44), which resides in the extracellular domain of the SSTR2 amino terminus, screened the peptide-bound MAbs by fluorescence microassay on D341 and D283 MED cells, and demonstrated homogeneous cell-surface binding, indicating that all cells expressed cell surface-detectable epitopes. Five radiolabeled MAbs were tested for immunoreactive fraction (IRF), affinity (KA) (Scatchard analysis vs. D341 MED cells), and internalization by MED cells. One IgG(3) MAb exhibited a 50-100% IRF, but low KA. Four IgG(2a) MAbs had 46-94% IRFs and modest KAs versus intact cells (0.21-1.2 x 10(8) M(-1)). Following binding of radiolabeled MAbs to D341 MED at 4 degrees C, no significant internalization was observed, which is consistent with results obtained in the absence of ligand. However, all MAbs exhibited long-term association with the cells; binding at 37 degrees C after 2 h was 65-66%, and after 24 h, 52-64%. In tests with MAbs C10 and H5, the number of cell surface receptors per cell, estimated by Scatchard and quantitative FACS analyses, was 3.9 x 10(4) for the "glial" phenotype DAOY MED cell line and 0.6-8.8 x 10(5) for four neuronal phenotype MED cell lines. Our results indicate a potential immunotherapeutic application for these MAbs. | |
| dc.description.version | Version of Record | |
| dc.identifier | ||
| dc.identifier.eissn | 1557-8348 | |
| dc.identifier.uri | ||
| dc.language | eng | |
| dc.language.iso | en_US | |
| dc.publisher | Mary Ann Liebert Inc | |
| dc.relation.ispartof | Hybridoma (Larchmt) | |
| dc.relation.isversionof | 10.1089/hyb.2009.0049 | |
| dc.relation.journal | Hybridoma | |
| dc.subject | Amino Acid Sequence | |
| dc.subject | Antibodies, Monoclonal | |
| dc.subject | Blotting, Western | |
| dc.subject | Cell Line, Tumor | |
| dc.subject | DNA Primers | |
| dc.subject | Enzyme-Linked Immunosorbent Assay | |
| dc.subject | Flow Cytometry | |
| dc.subject | Humans | |
| dc.subject | Immunohistochemistry | |
| dc.subject | Immunotherapy | |
| dc.subject | Medulloblastoma | |
| dc.subject | Protein Structure, Tertiary | |
| dc.subject | Receptors, Somatostatin | |
| dc.subject | Reverse Transcriptase Polymerase Chain Reaction | |
| dc.title | Detection of amino-terminal extracellular domain of somatostatin receptor 2 by specific monoclonal antibodies and quantification of receptor density in medulloblastoma. | |
| dc.type | Journal article | |
| duke.contributor.orcid | McLendon, Roger E|0000-0001-6682-4588 | |
| duke.contributor.orcid | Zalutsky, Michael R|0000-0002-5456-0324 | |
| duke.contributor.orcid | Bigner, Darell D|0000-0001-5548-4899 | |
| duke.date.pubdate | 2009-12-0 | |
| duke.description.issue | 6 | |
| duke.description.volume | 28 | |
| pubs.author-url | ||
| pubs.begin-page | 389 | |
| pubs.end-page | 403 | |
| pubs.issue | 6 | |
| pubs.organisational-group | Clinical Science Departments | |
| pubs.organisational-group | Duke | |
| pubs.organisational-group | Duke Cancer Institute | |
| pubs.organisational-group | Institutes and Centers | |
| pubs.organisational-group | Neurosurgery | |
| pubs.organisational-group | Pathology | |
| pubs.organisational-group | Radiation Oncology | |
| pubs.organisational-group | Radiology | |
| pubs.organisational-group | School of Medicine | |
| pubs.organisational-group | Surgery | |
| pubs.publication-status | Published | |
| pubs.volume | 28 |