| Quantifying multimodal contrast agent biological activity using near-infrared flow cytometry. | |
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MedLine Citation:
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PMID: 22539404 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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Prior to imaging agent use in preclinical studies and clinical diagnostics, biological activity must be validated. The Lindmo assay has been used conventionally to quantify radiolabeled antibody (Ab) immunoreactivity, although published findings suggest it does not provide consistently accurate results. We developed and tested a near-infrared (NIR) flow cytometry (FC) method for quantifying biological activity of a dual-labeled Ab for use as a multimodal contrast agent in small animal and human positron emission tomography and NIR fluorescence imaging. Antibody specific for epithelial cell adhesion molecule was conjugated to DOTA-NHS-ester, labeled with IRDye 800CW and further labeled with (64) Cu or nonradioactive Cu prior to reacting with human prostate cancer cells for testing by the Lindmo or FC method, respectively. Immunoreactivity of the dual-labeled agent was found to be 76.4 ± 15.7% by the Lindmo assay. When tested with and without Cu labeling using NIR FC, the biological activity was found to be 73.1 ± 7.7 and 79.4 ± 8.1%, respectively. No significant differences were found between these activity levels (p > 0.05), supporting NIR FC as an alternative method for measuring immunoreactivity and demonstrating that Cu labeling does not significantly affect the agent's ability to bind to its target. Biological activity was significantly reduced when the NIR dye-to-protein ratio was increased 3- to 4-fold in agent preparations when tested by FC and the Lindmo assay. In summary, NIR FC is an alternative with similar specificity and sensitivity, and greater reproducibility relative to the Lindmo assay for quantifying biological activity of NIR fluorophore-labeled, multimodal imaging agents. Copyright © 2012 John Wiley & Sons, Ltd. |
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Authors:
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Mary A Hall; Melissa B Aldrich; Ali Azhdarinia; Pier-Anne Lachance; Holly Robinson; Amy Hazen; David L Haviland; Eva M Sevick-Muraca |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Contrast media & molecular imaging Volume: 7 ISSN: 1555-4317 ISO Abbreviation: Contrast Media Mol Imaging Publication Date: 2012 May |
Date Detail:
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Created Date: 2012-04-27 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101286760 Medline TA: Contrast Media Mol Imaging Country: United States |
Other Details:
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Languages: eng Pagination: 338-45 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 John Wiley & Sons, Ltd. |
Affiliation:
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Center for Molecular Imaging, The Brown Foundation Institute of Molecular Medicine at The University of Texas Health Science Center, Houston, TX, 77030, USA. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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