Antibody Conjugate Assembly on Ultrasound-Confined Microcarrier Particles.
Antibody Conjugate Assembly on Ultrasound-Confined Microcarrier Particles.
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DOI：
10.1021/acsbiomaterials.0c01162
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发表时间：
2020-11-09
影响因子：
5.8
通讯作者：
Meacham JM
中科院分区：
文献类型：
--
作者：
Binkley MM;Cui M;Berezin MY;Meacham JM
生物缀合物是重要的下一代药物和显像剂。组装这些日益复杂的结构需要精确控制加工条件，这对传统的人工合成是一个挑战。这种不足促使人们寻求新的方法来有效地控制高分子量生物制剂如蛋白质、碳水化合物和核酸的修饰。我们报告了一种新型的，免提的，半自动化的平台，用于合成操作的生物分子使用声响应的微粒作为三维反应基板。微流体反应器结合了纵向声阱，其控制局部声场内的化学反应。由该场产生的力使微尺度基底抵抗参与的化学试剂的连续流动。因此，底物和反应物的运动是解耦的，使得能够精确控制多步反应条件，并以最小的用户输入提供高产率、高纯度的产物。我们通过将临床相关抗体与小分子结合来证明这些能力。珠上合成包括抗体的捕获、荧光标签的偶联、产物纯化和产物释放。通过使用流式细胞术测定的绿色（抗体）和红色（小分子染料）通道中值强度的49和11倍增加来确认荧光标记抗体的成功捕获和修饰。组装在声学响应的超声限制的微粒上的抗体缀合物表现出与由熟练技术人员手动制备的那些抗体缀合物相似的质量和数量。
Bioconjugates are important next-generation drugs and imaging agents. Assembly of these increasingly complex constructs requires precise control over processing conditions, which is a challenge for conventional manual synthesis. This inadequacy has motivated the pursuit of new approaches for efficient, controlled modification of high-molecular-weight biologics such as proteins, carbohydrates, and nucleic acids. We report a novel, hands-free, semiautomated platform for synthetic manipulation of biomolecules using acoustically responsive microparticles as three-dimensional reaction substrates. The microfluidic reactor incorporates a longitudinal acoustic trap that controls the chemical reactions within a localized acoustic field. Forces generated by this field immobilize the microscale substrates against the continuous flow of participating chemical reagents. Thus, the motion of substrates and reactants is decoupled, enabling exquisite control over multistep reaction conditions and providing high-yield, high-purity products with minimal user input. We demonstrate these capabilities by conjugating clinically relevant antibodies with a small molecule. The on-bead synthesis comprises capture of the antibody, coupling of a fluorescent tag, product purification, and product release. Successful capture and modification of a fluorescently labeled antibody are confirmed via fold increases of 49 and 11 in the green (antibody)- and red (small-molecule dye)-channel median intensities determined using flow cytometry. Antibody conjugates assembled on acoustically responsive, ultrasound-confined microparticles exhibit similar quality and quantity to those prepared manually by a skilled technician.
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