Linked Life Data

17508710

Source:http://linkedlifedata.com/resource/pubmed/id/17508710

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2007-7-19
pubmed:abstractText
We report a binary targeted enzymatic system that is composed of two covalent monoclonal antibody conjugates for specific labeling of cellular targets in vivo. The system utilizes low-molecular weight peroxidase-reducing substrates synthesized by linking 5-hydroxytryptamine (serotonin) with DTPA (5HT-DTPA) for magnetic resonance and radionuclide imaging or with Cy5.5 for near-infrared optical imaging. Initially, the conjugation reaction conditions were optimized to achieve a low level of antiepidermal growth factor receptor (EGFR) antibody (EMD 72000) modification with the N-hydroxysuccinimide ester of 4-hydrazinonicotinate acetone hydrazone (SANH), yielding mAb-HNH conjugate. The resultant modified antibodies were incubated with the periodate-oxidized peroxidase (HRP) or 4-formylbenzoyl-conjugated glucose oxidase (GO), followed by the purification of the resultant mAb-enzyme conjugates by size-exclusion HPLC. The conjugates were further characterized by electrophoresis and were tested by cross-titration on A431 EGFR+ squamous carcinoma or SW620 adenocarcinoma cells (negative control). The conjugates at the optimized concentration ratios were further tested using near-infrared fluorescence microscopy in the presence of Cy5.5 monocarboxy-5-hydroxytryptamide. Further in vitro experiments demonstrated that (1) antibody binding was specific and could be inhibited by free antibody; (2) both antibody conjugates exhibited high enzymatic activity after the binding to the cells; (3) 111In-labeled 5-HT-DTPA was avidly binding to EGFR-positive cells only if both HRP- and GO-conjugates were bound to the cells. The conjugates were tested in vivo using a SPECT imaging experiment, which demonstrated the accumulation of 111In-labeled 5-HT-DTPA substrate at the site containing both conjugates.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1043-1802
pubmed:author
pubmed:issnType
Print
pubmed:volume
18
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1123-30
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:17508710-Animals, pubmed-meshheading:17508710-Antibodies, Monoclonal, pubmed-meshheading:17508710-Carbocyanines, pubmed-meshheading:17508710-Catalysis, pubmed-meshheading:17508710-Cell Line, pubmed-meshheading:17508710-Fluorescent Dyes, pubmed-meshheading:17508710-Glucose Oxidase, pubmed-meshheading:17508710-Horseradish Peroxidase, pubmed-meshheading:17508710-Humans, pubmed-meshheading:17508710-Indium Radioisotopes, pubmed-meshheading:17508710-Magnetic Resonance Imaging, pubmed-meshheading:17508710-Mice, pubmed-meshheading:17508710-Microscopy, Fluorescence, pubmed-meshheading:17508710-Pentetic Acid, pubmed-meshheading:17508710-Receptor, Epidermal Growth Factor, pubmed-meshheading:17508710-Serotonin, pubmed-meshheading:17508710-Tomography, Emission-Computed, Single-Photon
pubmed:articleTitle
Synthesis and testing of a binary catalytic system for imaging of signal amplification in vivo.
pubmed:affiliation
Department of Radiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA. Alexei.Bogdanov@umassmed.edu
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural