本期文章：《自然—方法学》：Online/在线发表

香港中文大学Ho Ko和Hei Ming Lai合作近期取得重要工作进展，他们研究提出了用于热加速深度免疫染色的抗体稳定化方案。相关研究成果2022年9月1日在线发表于《自然—方法学》杂志上。

研究人员开发了一种简单、便于推广的化学方法，用于稳定成品抗体的热变性和化学变性。他们证明了稳定的抗体（称为SPEAR）可以承受长达4周的55°C连续高温和苛刻的变性剂，并将这种方法应用于33种经过测试的抗体。SPEARs能够灵活地应用热循环和变性剂，以动态调节它们的结合动力学、反应平衡、大分子扩散和聚集倾向。

研究人员特别表明，SPEARs 允许使用热促进的三维免疫标记策略（称为ThICK染色），在72小时内实现整个小鼠大脑的免疫标记，以及在人脑组织中以少三倍的抗体量进行近四倍的深度渗透免疫标记。凭借更快的深层组织免疫标记以及与组织处理和清除方法的广泛兼容性，无需任何专用设备，研究人员预计ThICK染色与SPEARs在深层免疫染色中将具有广泛适用性。

据了解，抗体由于其高反应特异性而具有多种不同的应用，但在需要更高工作温度时对变性敏感。

附：英文原文

Title: Antibody stabilization for thermally accelerated deep immunostaining

Author: Lai, Hei Ming, Tang, Yumi, Lau, Zachary Y. H., Campbell, Robert A. A., Yau, Juno C. N., Chan, Caleb C. Y., Chan, Danny C. W., Wong, Tin Yan, Wong, Harriet K. T., Yan, Leo Y. C., Wu, William K. K., Wong, Sunny H., Kwok, Ka-Wai, Wing, Yun-Kwok, Lam, Henry H. N., Ng, Ho-Keung, Mrsic-Flogel, Thomas D., Mok, Vincent C. T., Chan, Jason Y. K., Ko, Ho

Issue&Volume: 2022-09-01

Abstract: Antibodies have diverse applications due to their high reaction specificities but are sensitive to denaturation when a higher working temperature is required. We have developed a simple, highly scalable and generalizable chemical approach for stabilizing off-the-shelf antibodies against thermal and chemical denaturation. We demonstrate that the stabilized antibodies (termed SPEARs) can withstand up to 4weeks of continuous heating at 55°C and harsh denaturants, and apply our method to 33 tested antibodies. SPEARs enable flexible applications of thermocycling and denaturants to dynamically modulate their binding kinetics, reaction equilibrium, macromolecular diffusivity and aggregation propensity. In particular, we show that SPEARs permit the use of a thermally facilitated three-dimensional immunolabeling strategy (termed ThICK staining), achieving whole mouse brain immunolabeling within 72h, as well as nearly fourfold deeper penetration with threefold less antibodies in human brain tissue. With faster deep-tissue immunolabeling and broad compatibility with tissue processing and clearing methods without the need for any specialized equipment, we anticipate the wide applicability of ThICK staining with SPEARs for deep immunostaining.