The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Delivery of Drugs and Genes Group (D2G2), St Lucia, QLD 4072, Australia.
ACS Appl Mater Interfaces. 2012 May;4(5):2483-9. doi: 10.1021/am3001727. Epub 2012 Mar 20.
New technologies are needed to translate biomarker discovery research into simple, inexpensive, and effective molecular diagnostic assays for use by clinicians or patients to guide and monitor treatment. Microprojection arrays were recently introduced as tools which, when applied to the skin, penetrate into the dermal tissue, and capture specific circulating biomarkers. In our initial work on Microprojection arrays, carbodiimide chemistry was used to immobilize biomarker-specific probes for affinity capture in vivo using a mouse model. However, as the observed capture efficiencies were relatively low, with significant variation across the surface, here we investigated the surface modifications to (a) determine the source of the variability and (b) find ways of improving capture efficiency. We found the protein immobilization step accounted for almost all of the variability in surface uniformity. Varying the protein immobilization conditions following a standard carbodiimide activation process resulted in a reduction in overall variation 14-fold and an increase in captured biomarker amount ∼18-fold. In conclusion, we found that investigating and optimizing the surface chemistry of microprojection array devices led to drastic improvements in capturing biomarkers from skin fluid.
需要新技术将生物标志物发现研究转化为简单、廉价且有效的分子诊断检测方法,以便临床医生或患者用于指导和监测治疗。微喷射阵列最近被引入作为工具,当应用于皮肤时,它们会穿透真皮组织,并捕获特定的循环生物标志物。在我们对微喷射阵列的初步研究中,使用碳二亚胺化学将生物标志物特异性探针固定在体内,用于使用小鼠模型进行亲和捕获。然而,由于观察到的捕获效率相对较低,并且表面存在很大差异,因此我们研究了表面修饰,以确定(a)变异性的来源和(b)提高捕获效率的方法。我们发现蛋白质固定化步骤几乎占表面均匀性变化的全部。在标准的碳二亚胺活化过程之后改变蛋白质固定化条件,可使整体变化降低 14 倍,捕获的生物标志物数量增加约 18 倍。总之,我们发现研究和优化微喷射阵列器件的表面化学可显著提高从皮肤液中捕获生物标志物的效率。