We describe a novel analytical method for rapid and robust identification, mapping and relative quantitation of glycopeptides from SARS-CoV-2 Spike protein. The method may be executed using any LC-TOF mass spectrometer, requires no specialised knowledge of glycan analysis and makes use of the differential resolving power of reversed phase HPLC. While this separation resolves peptides with high efficiency, glycans are resolved poorly if at all. Consequently, glycopeptides consisting of the same peptide bearing different glycan structures will all possess very similar retention times and co-elute. While this has previously been viewed as a disadvantage, we show that shared retention time can be used to map multiple glycan species to the same peptide and location. In combination with MSMS and pseudo MS3, we have constructed a detailed mass-retention time database for Spike. This database allows any ESI-TOF equipped lab to reliably identify and quantify spike glycans from a single overnight elastase protein digest in less than 90 minutes.

中文翻译：

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质量保留时间指纹图谱对SARS-CoV-2穗糖肽的鉴定，定位和相对定量

我们描述了一种新的分析方法，可用于快速，可靠地识别，定位和绘制SARS-CoV-2 Spike蛋白糖肽的相对定量。该方法可以使用任何LC-TOF质谱仪执行，不需要专门的聚糖分析知识，并利用反相HPLC的差分分辨能力。尽管这种分离可以高效分离肽，但聚糖分离效果却很差。因此，由具有不同聚糖结构的相同肽组成的糖肽都将具有非常相似的保留时间并共洗脱。尽管以前认为这是一个缺点，但我们显示共享的保留时间可用于将多个聚糖种类映射到相同的肽和位置。结合MSMS和伪MS3，我们已经为Spike建立了详细的质量保留时间数据库。该数据库使任何配备ESI-TOF的实验室都能在不到90分钟的时间内，从单一的过夜弹性蛋白酶蛋白质消化物中可靠地鉴定和定量加标的聚糖。