Centro de Astrobiología (INTA-CSIC), Madrid, Spain.
Astrobiology. 2011 Jan-Feb;11(1):15-28. doi: 10.1089/ast.2010.0501. Epub 2011 Feb 6.
The search for unequivocal signs of life on other planetary bodies is one of the major challenges for astrobiology. The failure to detect organic molecules on the surface of Mars by measuring volatile compounds after sample heating, together with the new knowledge of martian soil chemistry, has prompted the astrobiological community to develop new methods and technologies. Based on protein microarray technology, we have designed and built a series of instruments called SOLID (for "Signs Of LIfe Detector") for automatic in situ detection and identification of substances or analytes from liquid and solid samples (soil, sediments, or powder). Here, we present the SOLID3 instrument, which is able to perform both sandwich and competitive immunoassays and consists of two separate functional units: a Sample Preparation Unit (SPU) for 10 different extractions by ultrasonication and a Sample Analysis Unit (SAU) for fluorescent immunoassays. The SAU consists of five different flow cells, with an antibody microarray in each one (2000 spots). It is also equipped with an exclusive optical package and a charge-coupled device (CCD) for fluorescent detection. We demonstrated the performance of SOLID3 in the detection of a broad range of molecular-sized compounds, which range from peptides and proteins to whole cells and spores, with sensitivities at 1-2 ppb (ng mL⁻¹) for biomolecules and 10⁴ to 10³ spores per milliliter. We report its application in the detection of acidophilic microorganisms in the Río Tinto Mars analogue and report the absence of substantial negative effects on the immunoassay in the presence of 50 mM perchlorate (20 times higher than that found at the Phoenix landing site). Our SOLID instrument concept is an excellent option with which to detect biomolecules because it avoids the high-temperature treatments that may destroy organic matter in the presence of martian oxidants.
在其他行星体上寻找明确的生命迹象是天体生物学的主要挑战之一。在对火星表面样品进行加热后,通过测量挥发性化合物来探测有机分子的尝试均以失败告终,再加上对火星土壤化学的新认识,促使天体生物学界开发新的方法和技术。基于蛋白质微阵列技术,我们设计并制造了一系列名为 SOLID(代表“生命迹象探测器”)的仪器,用于自动原位检测和识别来自液体和固体样品(土壤、沉积物或粉末)中的物质或分析物。在这里,我们展示了 SOLID3 仪器,它能够执行夹心和竞争免疫测定,由两个独立的功能单元组成:一个用于 10 种不同超声提取的样品制备单元 (SPU) 和一个用于荧光免疫测定的样品分析单元 (SAU)。SAU 由五个不同的流动池组成,每个流动池中都有一个抗体微阵列(共 2000 个斑点)。它还配备了专用的光学组件和电荷耦合器件 (CCD) 用于荧光检测。我们展示了 SOLID3 在检测广泛的分子大小化合物方面的性能,这些化合物的范围从肽和蛋白质到整个细胞和孢子,对于生物分子的灵敏度为 1-2 ppb(ng mL⁻¹),对于每毫升 10⁴ 到 10³ 个孢子。我们报告了它在检测 Rio Tinto Mars 模拟物中嗜酸微生物中的应用,并报告了在存在 50 mM 高氯酸盐(比凤凰着陆点高 20 倍)的情况下,对免疫测定没有实质性的负面影响。我们的 SOLID 仪器概念是检测生物分子的绝佳选择,因为它避免了在存在火星氧化剂的情况下可能破坏有机物的高温处理。