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Synthetically Modified Viral Capsids as Versatile Carriers for Use in Antibody-Based Cell Targeting

机译:合成修饰的病毒衣壳作为多功能载体用于基于抗体的细胞靶向

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摘要

The present study describes an efficient and reliable method for the preparation of MS2 viral capsids that are synthetically modified with antibodies using a rapid oxidative coupling strategy. The overall protocol delivers conjugates in high yields and recoveries, requires a minimal excess of antibody to achieve modification of more than 95% of capsids, and can be completed in a short period of time. Antibody–capsid conjugates targeting extracellular receptors on human breast cancer cell lines were prepared and characterized. Notably, conjugation to the capsid did not significantly perturb the binding of the antibodies, as indicated by binding affinities similar to those obtained for the parent antibodies. An array of conjugates was synthesized with various reporters on the interior surface of the capsids to be used in cell studies, including fluorescence-based flow cytometry, confocal microscopy, and mass cytometry. The results of these studies lay the foundation for further exploration of these constructs in the context of clinically relevant applications, including drug delivery and in vivo diagnostics.
机译:本研究描述了一种有效且可靠的方法,用于制备通过快速氧化偶联策略经抗体合成修饰的MS2病毒衣壳。整个方案以高收率和高回收率提供结合物,仅需极少量的抗体即可修饰超过95%的衣壳,并且可以在短时间内完成。制备并表征了靶向人乳腺癌细胞系上细胞外受体的衣壳结合物。值得注意的是,与衣壳的缀合不会显着干扰抗体的结合,如与亲本抗体获得的结合亲和力相似的结合亲和力所表明的。在衣壳的内表面上用各种报道分子合成了一系列缀合物,用于细胞研究,包括基于荧光的流式细胞术,共聚焦显微镜和大规模细胞术。这些研究的结果为在临床相关应用(包括药物输送和体内诊断)中进一步探索这些构建体奠定了基础。

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