Assessment of Free Dye in Solutions of Dual-Labeled Antibody Conjugates for In Vivo Molecular Imaging
Assessment of Free Dye in Solutions of Dual-Labeled Antibody Conjugates for In Vivo Molecular Imaging
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DOI：
10.1007/s11307-010-0328-7
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发表时间：
2011-02-01
影响因子：
3.1
通讯作者：
Sevick-Muraca, Eva M.
中科院分区：
文献类型：
--
作者：
Aldrich, Melissa B.;Wang, XueJuan;Sevick-Muraca, Eva M.
Recent preclinical and clinical studies show that dyes that excite and fluoresce in the near-infrared range may be used for tracking and detecting disease targets in vivo. A method for quantifying free dye molecules in antibody conjugate preparations is required for agent batch release and for translation into the clinic.Herein, we developed and validated a SDS-PAGE method to determine the percentage of free IRDye 800CW in (DTPA)(n)-trastuzumab-(IRDye 800)(m) conjugate sample preparations in which high-performance liquid chromatography (HPLC) assessment of free dye was not possible.The SDS-PAGE assay was accurate and valid for free IRDye 800CW amounts between 38 and 4 mol% of total dye. Gel sample preparation reagent affected the specificity of the assay, and lower and upper limits of quantitation and detection were determined.This method may be applicable to other near-infrared dye-conjugated antibody-based imaging agents in which HPLC assessment of purity is not feasible. This validated method for quality assurance will facilitate the translation of dual-labeled antibody conjugates for nuclear and optical imaging.
