Lithographic Patterning of Nanoscale Arrays of the Oxidase Enzyme CotA: Effects on Activity and Stability
Lithographic Patterning of Nanoscale Arrays of the Oxidase Enzyme CotA: Effects on Activity and Stability
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DOI：
10.1002/admt.202200490
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发表时间：
2022-07
影响因子：
6.8
通讯作者：
Silvia Fruncillo;Y. Toh;C. Blanford;X. Su;Hong Liu;L. Wong
中科院分区：
文献类型：
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作者：
Silvia Fruncillo;Y. Toh;C. Blanford;X. Su;Hong Liu;L. Wong
这项研究比较了由两种光刻方法产生的带有氧化酶科塔的纳米级阵列的酶活性：聚合物笔光刻（PPL），一种用于小面积制造的扫描探针光刻（100 cm 2）;和步进光刻，一种用于微电子领域的大规模方法（>300 cm 2）。在这两种情况下，600 nm的金特征阵列都是通过生物偶联方法产生的，并通过科塔进行功能化，从而提供均匀的蛋白质取向。然后在100天内定量这些阵列的酶活性。通过光刻法生产的酶阵列在制造后立即具有更高的氧化活性，但与PPL生产的酶阵列相比，降解更快。这一结果是由于光刻法生产的阵列体表面钝化较差，导致大量非特异性吸附的酶。然而，一旦调整结果以考虑钝化和表面积的差异，就发现固定在金特征上的酶基本上具有相同的活性，而与所使用的平版印刷方法无关。因此，这些结果表明，PPL是一种合适的方法，用于在放大之前对生物设备进行原型设计，前提是适当考虑制造特征的设计。
This study compares the enzymatic activity of nanoscale arrays bearing the oxidase CotA that are produced by two lithographic methods: polymer pen lithography (PPL), a scanning probe lithography for small‐area fabrication (≈1 cm2); and stepper photolithography, a large‐scale method (>300 cm2) used in the microelectronics sector. In both cases, arrays of 600 nm gold features are produced and functionalized with CotA through a bioconjugation method that gives uniform protein orientation. The enzyme activity of these arrays is then quantified over 100 days. The enzyme arrays produced by photolithography give higher oxidation activities immediately after fabrication but degrade more rapidly when compared to those produced by PPL. This result is due to the poorer passivation on the bulk surface of photolithographically produced arrays, resulting in a greater amount of non‐specifically adsorbed enzymes. However, once the results are adjusted to account for the differences in passivation and surface area, it is found that the enzymes immobilized on the gold features give essentially the same activity regardless of the lithographic method used. Thus, these results suggest PPL is a suitable method for prototyping biodevices prior to scale‐up, provided that due consideration is given to the design of the fabricated features.