Department of Biomedical Engineering, Medical School, Shenzhen University, Shenzhen, 518060, China.
Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, Department of Biomedical Engineering, Medical School, Shenzhen University, Shenzhen, 518060, China.
Adv Biol (Weinh). 2023 Aug;7(8):e2300111. doi: 10.1002/adbi.202300111. Epub 2023 May 13.
Antibody arrays have great implications in many biomedical settings. However, commonly used patterning methods have difficulties in generating antibody arrays with both high resolution and multiplexity, limiting their applications. Here, a convenient and versatile technique for the patterning of multiple antibodies with resolution down to 20 µm is reported using micropillar-focused droplet printing and microcontact printing. Droplets of antibody solutions are first printed and stably confined on the micropillars of a stamp, and then the antibodies absorbed on the micropillars are contact-printed to the target substrate, generating antibody patterns faithfully replicating the micropillar array. The effect of different parameters on the patterning results is investigated, including hydrophobicity of the stamps, override time of the droplet printing, incubation time, and the diameters of the capillary tips and micropillars. To demonstrate the utility of the method, multiplex arrays of anti-EpCAM and anti-CD68 antibodies is generated to capture breast cancer cells and macrophages, respectively, on the same substrate, and successful capturing of individual cell types and enrichment among the cells are achieved. It is envision that this method would serve as a versatile and useful protein patterning tool for biomedical applications.
抗体阵列在许多生物医学环境中具有重要意义。然而，常用的图案化方法在生成具有高分辨率和多重性的抗体阵列方面存在困难，限制了它们的应用。在这里，报道了一种使用微柱聚焦液滴打印和微接触印刷来对分辨率低至 20 µm 的多种抗体进行图案化的简便、通用的技术。首先打印抗体溶液的液滴并将其稳定地限制在印章的微柱上，然后将吸附在微柱上的抗体接触印刷到目标基底上，从而生成忠实地复制微柱阵列的抗体图案。研究了不同参数对图案化结果的影响，包括印章的疏水性、液滴打印的超越时间、孵育时间以及毛细尖端和微柱的直径。为了证明该方法的实用性，生成了抗 EpCAM 和抗 CD68 抗体的多重阵列，分别在同一基底上捕获乳腺癌细胞和巨噬细胞，并成功捕获了单个细胞类型和细胞之间的富集。可以预见，该方法将成为生物医学应用中一种通用且有用的蛋白质图案化工具。
