用于体内分子成像的双标记抗体缀合物溶液中游离染料的评估。
Assessment of free dye in solutions of dual-labeled antibody conjugates for in vivo molecular imaging.
机构信息
Division of Molecular Imaging, The Brown Foundation Institute of Molecular Medicine, University of Texas Health Sciences Center-Houston, Houston, TX, USA.
出版信息
Mol Imaging Biol. 2011 Feb;13(1):32-42. doi: 10.1007/s11307-010-0328-7.
PURPOSE
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.
PROCEDURES
Herein, we developed and validated a SDS-PAGE method to determine the percentage of free IRDye 800 CW in (DTPA)(n)-trastuzumab-(IRDye 800)(m) conjugate sample preparations in which high-performance liquid chromatography (HPLC) assessment of free dye was not possible.
RESULTS
The SDS-PAGE assay was accurate and valid for free IRDye 800 CW 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.
CONCLUSION
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.
目的
最近的临床前和临床研究表明，近红外激发和荧光染料可用于体内跟踪和检测疾病靶标。对于试剂批次放行和转化为临床应用，需要一种定量抗体缀合物制剂中游离染料分子的方法。
方法
本文开发并验证了 SDS-PAGE 方法，用于测定（DTPA）（n）-曲妥珠单抗-（IRDye 800）（m）缀合物样品制剂中游离 IRDye 800CW 的百分比，其中高效液相色谱（HPLC）无法评估游离染料。
结果
SDS-PAGE 测定法对游离 IRDye 800CW 量为总染料的 38%至 4mol%之间是准确和有效的。凝胶样品制备试剂会影响测定的特异性，并且确定了定量和检测的下限和上限。
结论
该方法可能适用于其他近红外染料标记的基于抗体的成像剂，其中无法进行 HPLC 纯度评估。这种经过验证的质量保证方法将促进用于核和光学成像的双标记抗体缀合物的转化。
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