图像细胞术测定抗体滴度

R. Hertzog, D. Popescu, Bianca Palade
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引用次数: 0

摘要

“为了防止病毒感染的传播或评估疫苗接种的有效性,迫切需要快速确定和描述可能的治疗方案的方法。血清学方法,通常用于抗体滴定,信息丰富,但提供的数据有时是有限的。成像细胞术可以是一种有效的方法来表征潜在的治疗性抗体,以对抗病毒感染。采用基于BIOCHIP技术的间接免疫荧光法检测抗黄热病病毒IgG,通过稀释阳性对照血清和接种黄热病疫苗的人的血液样本,实现了校准曲线。然后通过图像细胞术对获得的图像进行分析,其中包括:图像预处理,去除被认为最具代表性的细胞核进行测量,间接测量每个代表性细胞的特定校正总细胞荧光(CTCF)的五倍,并计算平均CTCF值。我们计算了每种情况下的平均CTCF值,并根据制造商的建议将CTCF值与抗体滴度相关联。图像细胞术能够快速确定抗体与宿主细胞的直接结合,并可应用于研究其他病原体-抗体相互作用,从而影响未来对病毒病原体的研究。”
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ANTIBODY TITER ASSESSMENT BY IMAGE CYTOMETRY
"In order to prevent the spread of viral infections or to assess the effectiveness of vaccination, there is an urgent need for methods to quickly identify and characterize possible treatment options. The serological methods, commonly used for antibody titration, are informative yet the data provided are sometimes limited. Imaging cytometry can be an effective approach for characterizing potential therapeutic antibodies to combat viral infections. Using an indirect immunofluorescence test, based on BIOCHIP technology to detect anti-yellow fever virus IgG, we realized a calibration curve based on dilution of a positive control serum and a blood sample obtained from a person who has received the yellow fever vaccine. The obtained images were then analysed by image cytometry which involved: image pre-processing, removal of the cell nucleus considered to be the most representative for measurement, indirect measurement of five times the specific corrected total cellular fluorescence (CTCF) for each representative cell and calculation of the average CTCF value. We calculated the mean CTCF values for each condition and correlated the CTCF value with the antibody titer, according to the manufacturer's recommendations. Image cytometry has the ability to rapidly determine the direct binding of antibodies to host cells and can be applied to study other pathogen-antibody interactions, thus impacting future research on viral pathogens. "
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