用于通过液相色谱-质谱联用分析抗菌肽Kn2-7/dKn2-7在人血清中稳定性的优化肽提取方法。
Optimized peptide extraction method for analysis of antimicrobial peptide Kn2-7/dKn2-7 stability in human serum by LC-MS.
作者信息
Chen Wen, Kirui Dickson, Millenbaugh Nancy J
机构信息
Craniofacial Health & Restorative Medicine, Naval Medical Research Unit San Antonio, 3650 Chambers Pass, Building 3610, Joint Base San Antonio-Fort Sam Houston, TX 78234, USA.
出版信息
Future Sci OA. 2022 Jul 20;8(6):FSO807. doi: 10.2144/fsoa-2022-0013. eCollection 2022 Jul.
AIM
To develop an extraction protocol and determine stability for antimicrobial peptide (AMP) Kn2-7 and its d-enantiomer dKn2-7 in human serum.
MATERIALS & METHODS: We compared use of ethanol, acetonitrile, RapiGest SF Surfactant and 1% formic acid in ethanol for AMP recovery from serum prior to liquid chromatography-mass spectrometry quantification.
RESULTS
Precipitation of samples with 1% formic acid in ethanol caused the least amount of AMP loss during extraction from serum. Time-course experiments revealed dKn2-7 was significantly more stable than Kn2-7 in 25% serum, with 78.5% of dKn2-7 and only 1.0% of Kn2-7 remaining after 24 h at 37°C.
CONCLUSION
The optimized method significantly increased peptide recovery and allowed more accurate and consistent quantification of Kn2-7 and dKn2-7 serum stability.
目的
制定一种提取方案，并确定抗菌肽（AMP）Kn2-7及其d-对映体dKn2-7在人血清中的稳定性。
材料与方法
在进行液相色谱-质谱定量分析之前，我们比较了使用乙醇、乙腈、RapiGest SF表面活性剂和乙醇中的1%甲酸从血清中回收AMP的情况。
结果
用乙醇中的1%甲酸沉淀样品在从血清中提取过程中导致的AMP损失量最少。时间进程实验表明，在25%血清中，dKn2-7比Kn2-7稳定得多，在37°C下24小时后，dKn2-7剩余78.5%，而Kn2-7仅剩余1.0%。
结论
优化后的方法显著提高了肽的回收率，并能更准确、一致地定量Kn2-7和dKn2-7的血清稳定性。
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