Evaluation of the Binding Preference of Microtubes for Nanoproteomics Sample Preparation.
Evaluation of the Binding Preference of Microtubes for Nanoproteomics Sample Preparation.
复制标题
DOI：
10.1021/acs.jproteome.2c00477
复制
发表时间：
2023-01-06
影响因子：
4.4
通讯作者：
Gao, Yu
中科院分区：
文献类型：
--
作者：
Zhang, Ziwei;Gao, Yu
在大规模蛋白质组学样品制备中，蛋白质与容器之间的非特异性结合是一个经常被忽视的样品丢失原因。在纳米蛋白质组学中，由于样本量小，这种吸收损失不再是可以忽略的，研究人员经常采用低结合塑料器皿来最大限度地减少样本损失。然而，在科学文献中，很少有人讨论微管在减少蛋白质/肽结合方面的性能差异。因此，样品制备过程中样品丢失的确切影响还不是很清楚。在这里，我们研究了在纳米蛋白质组实验过程中蛋白质/多肽的损失。结果表明，不同微管在非特异性结合方面存在显著差异，不同微管的蛋白质回收率在10%以下到90%以上。有趣的是，我们发现储存温度也可能是从塑料容器中回收蛋白质的关键因素之一。此外，我们还根据识别出的蛋白质和多肽的物理特性，如等电点、疏水性、长度和电荷，研究了不同微管的结合偏好。我们的发现有助于更好地了解蛋白质组学样品制备过程中蛋白质/多肽的损失，并为研究人员选择合适的容器保存宝贵的样品提供进一步的指导。
Non-specific binding between the protein and the container is an often-neglected cause of sample loss in large-scale proteomics sample preparation. In nanoproteomics, due to the small sample size, this absorption loss is no longer negligible, and researchers often adopt low binding plasticware to minimize the sample loss. However, there has been little discussion in the scientific literature on the differences in microtube performance in reducing protein/peptide binding. Therefore, the exact impact of sample loss during the sample preparation is not well understood. Here, we investigated the protein/peptide loss during the nanoproteomics experiment process. Our results showed that there are significant differences in non-specific binding among tested microtubes, with a protein recovery rate ranging from less than 10% to over 90% for different microtubes. Interestingly, we found that storage temperature could also be one of the key factors that contribute to protein recovery from the plastic container. In addition, we investigated the binding preferences of different microtubes by the physical characteristics of identified proteins and peptides, such as isoelectric point, hydrophobicity, length, and charge. Our findings help to better understand protein/peptide loss in proteomics sample preparation and provide further guidance for researchers in choosing proper containers for their precious sample.
登录
查看更多内容
影响因子：
14.9
作者：
Perez-Riverol Y;Bai J;Bandla C;García-Seisdedos D;Hewapathirana S;Kamatchinathan S;Kundu DJ;Prakash A;Frericks-Zipper A;Eisenacher M;Walzer M;Wang S;Brazma A;Vizcaíno JA
通讯作者：
Vizcaíno JA
影响因子：
6.1
作者：
Larsen, Esben Kjaer Unmack;Larsen, Niels B.
通讯作者：
Larsen, Niels B.
影响因子：
2.5
作者：
Weikart, Christopher M.;Breeland, Adam P.;Maurer, Brian R.
通讯作者：
Maurer, Brian R.
影响因子：
2.9
作者：
SMITH, JA;HURRELL, JGR;LEACH, SJ
通讯作者：
LEACH, SJ
影响因子：
15.6
作者：
Rabe, Michael;Verdes, Dorinel;Seeger, Stefan
通讯作者：
Seeger, Stefan