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Antibody Conjugate Assembly on Ultrasound-Confined Microcarrier Particles
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-09-25 , DOI: 10.1021/acsbiomaterials.0c01162 Michael M Binkley 1 , Mingyang Cui 1 , Mikhail Y Berezin 1 , J Mark Meacham 1
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-09-25 , DOI: 10.1021/acsbiomaterials.0c01162 Michael M Binkley 1 , Mingyang Cui 1 , Mikhail Y Berezin 1 , J Mark Meacham 1
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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.
中文翻译：
超声封闭微载体颗粒上的抗体偶联物组装
生物偶联物是重要的下一代药物和显像剂。这些日益复杂的结构的组装需要对加工条件进行精确控制，这对传统的手动合成来说是一个挑战。这种不足促使人们寻求对高分子量生物制剂（如蛋白质、碳水化合物和核酸）进行有效、可控修饰的新方法。我们报告了一种新颖的、免提的、半自动化的平台，用于使用声学响应微粒作为三维反应底物对生物分子进行合成操作。微流体反应器包含一个纵向声波阱，可控制局部声场内的化学反应。由该场产生的力将微型基材固定在参与化学试剂的连续流动中。因此，底物和反应物的运动是分离的，能够精确控制多步反应条件，并以最少的用户输入提供高产率、高纯度的产品。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。荧光标记抗体的成功捕获和修饰通过使用流式细胞术确定的绿色（抗体）和红色（小分子染料）通道中值强度增加 49 倍和 11 倍来确认。抗体偶联物组装在声学响应上，
更新日期：2020-11-09
中文翻译：
超声封闭微载体颗粒上的抗体偶联物组装
生物偶联物是重要的下一代药物和显像剂。这些日益复杂的结构的组装需要对加工条件进行精确控制，这对传统的手动合成来说是一个挑战。这种不足促使人们寻求对高分子量生物制剂（如蛋白质、碳水化合物和核酸）进行有效、可控修饰的新方法。我们报告了一种新颖的、免提的、半自动化的平台，用于使用声学响应微粒作为三维反应底物对生物分子进行合成操作。微流体反应器包含一个纵向声波阱，可控制局部声场内的化学反应。由该场产生的力将微型基材固定在参与化学试剂的连续流动中。因此，底物和反应物的运动是分离的，能够精确控制多步反应条件，并以最少的用户输入提供高产率、高纯度的产品。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。荧光标记抗体的成功捕获和修饰通过使用流式细胞术确定的绿色（抗体）和红色（小分子染料）通道中值强度增加 49 倍和 11 倍来确认。抗体偶联物组装在声学响应上，
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