School of Chemical Engineering, University of Campinas, Avenida Albert Einstein 500, 13083-852 Campinas, SP, Brazil.
Chemical Engineering Department, Massachusetts Institute of Technology, 77 Cambridge Avenue, Cambridge, Massachusetts 02139, United States.
Biomacromolecules. 2023 Jul 10;24(7):3159-3170. doi: 10.1021/acs.biomac.3c00224. Epub 2023 Jun 22.
The self-assembled layer-by-layer technique has attracted a great deal of attention as a method for engineering bio-functional surfaces under mild chemical conditions. The production of multilayer films, starting from newly designed building blocks, may be laborious, considering the inherent limitations for anticipating how minimal changes in the macromolecular composition may impact both film deposition and performance. This paper presents an automated, high-throughput approach to depositing polyelectrolyte multilayers (PEMs) in multiwell plates, enabling the screening of nearly 100 film formulations in the same process. This high-throughput layer-by-layer (HT-LbL) method runs in an affordable, fully commercial platform using Python-coded routines that can be easily adapted for the materials science lab settings. The HT-LbL system was validated by investigating the deposition of polysaccharide-based films in multiwell plates, probing the absorbance signal of ionically stained polyelectrolyte multilayers (PEMs) prepared in one single batch. The HT-LbL method was also used to investigate the deposition of PEMs with a small library of genetically engineered elastin-like polypeptides (ELPs) with different levels of ionizable and hydrophobic amino acid residues. The deposition of ELP/chitosan films was assessed based on the signal of fluorescently labeled species (chitosan or ELP-mCherry), demonstrating that both electrostatic and hydrophobic residues are essential for film buildup. The growth and surface properties of ELP-mCherry/chitosan films also seemed susceptible to the assembly pH, forming a higher film growth and a rougher and more hydrophobic surface for both polyelectrolytes deposited under a low ionization degree. Overall, this study illustrates the challenge of predicting the growth and properties of multilayer films and how the HT-LbL can accelerate the development of multilayer films that demand high levels of testing and optimization.
自组装层层技术作为一种在温和化学条件下工程生物功能表面的方法引起了极大的关注。从新设计的构建块开始生产多层膜，考虑到内在的局限性，即难以预测大分子组成的微小变化如何影响膜的沉积和性能，这可能是一项费力的工作。本文提出了一种在多孔板中自动、高通量沉积聚电解质多层（PEM）的方法，能够在同一过程中筛选近 100 种膜配方。这种高通量层层（HT-LbL）方法在一个经济实惠、完全商业化的平台上运行，使用 Python 编码例程，这些例程可以轻松适应材料科学实验室的设置。通过在多孔板中沉积多糖基膜，探测离子染色的聚电解质多层（PEM）在单个批次中的吸收信号，验证了 HT-LbL 系统。还使用 HT-LbL 方法研究了具有不同可离子化和疏水性氨基酸残基水平的基因工程弹性蛋白样多肽（ELP）的小文库的 PEM 沉积。基于荧光标记的物质（壳聚糖或 ELP-mCherry）的信号评估 ELP/壳聚糖膜的沉积，证明静电和疏水性残基对于膜的构建都是必不可少的。ELP-mCherry/壳聚糖膜的生长和表面性质似乎也容易受到组装 pH 值的影响，在低离子化程度下沉积的两种聚电解质形成了更高的膜生长和更粗糙、更疏水的表面。总的来说，这项研究说明了预测多层膜的生长和性能的挑战，以及 HT-LbL 如何加速需要高水平测试和优化的多层膜的开发。
