Progranulin Adsorbs to Polypropylene Tubes and Disrupts Functional Assays: Implications for Research, Biomarker Studies, and Therapeutics.
Progranulin Adsorbs to Polypropylene Tubes and Disrupts Functional Assays: Implications for Research, Biomarker Studies, and Therapeutics.
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DOI：
10.3389/fnins.2020.602235
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发表时间：
2020
影响因子：
4.3
通讯作者：
Kao AW
中科院分区：
文献类型：
--
作者：
Gururaj S;Sampognaro PJ;Argouarch AR;Kao AW
颗粒蛋白前体（PGRN）是一种受严格调控的分泌型糖蛋白，其参与广泛的生物过程，由于其参与肿瘤、神经发育和神经退行性疾病而引起科学界的极大兴趣。特别是，颗粒蛋白前体单倍不足导致额颞叶痴呆。在用HIS标记的重组人（rh）PGRN蛋白进行实验时，我们观察到由于其吸附到聚丙烯（PPE）微量离心管上，蛋白从溶液中的可测量的消耗。在这项研究中，我们已经量化了rhPGRN吸附PPE管的程度，而不同的实验条件，包括孵育时间和温度。我们发现，即使在很短的孵育期后，也有约25-35%的rhPGRN被吸附到PPE管的表面。然后，我们直接显示了PGRN吸附在功能测定中的有害影响，并推荐了替代实验室器皿以最大限度地减少这些影响。虽然一些纯化的蛋白质和肽吸附到聚合物塑料的风险已被表征，这是第一次报告的rhPGRN吸附。此外，由于PGRN目前正在基础科学实验室进行体外研究和转化实验室研究，以调查PGRN作为定量痴呆生物标志物和潜在的替代疗法，因此这里报告的观察结果具有广泛的影响力，可能会显著影响未来以颗粒蛋白前体生物学为中心的实验的设计和解释。
Progranulin (PGRN) is a tightly regulated, secreted glycoprotein involved in a wide range of biological processes that is of tremendous interest to the scientific community due to its involvement in neoplastic, neurodevelopmental, and neurodegenerative diseases. In particular, progranulin haploinsufficiency leads to frontotemporal dementia. While performing experiments with a HIS-tagged recombinant human (rh) PGRN protein, we observed a measurable depletion of protein from solution due to its adsorption onto polypropylene (PPE) microcentrifuge tubes. In this study, we have quantified the extent of rhPGRN adsorption to PPE tubes while varying experimental conditions, including incubation time and temperature. We found that ∼25–35% of rhPGRN becomes adsorbed to the surface of PPE tubes even after a short incubation period. We then directly showed the deleterious impact of PGRN adsorption in functional assays and have recommended alternative labware to minimize these effects. Although the risk of adsorption of some purified proteins and peptides to polymer plastics has been characterized previously, this is the first report of rhPGRN adsorption. Moreover, since PGRN is currently being studied and utilized in both basic science laboratories to perform in vitro studies and translational laboratories to survey PGRN as a quantitative dementia biomarker and potential replacement therapy, the reported observations here are broadly impactful and will likely significantly affect the design and interpretation of future experiments centered on progranulin biology.
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影响因子：
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