查找使用阿拉丁产品的科研文献

Biofunctionalized dissolvable hydrogel microbeads enable efficient characterization of native protein complexes

Shao Xinyang,Tian Meng,Yin Junlong,Duan Haifeng,Tian Ye,Wang Hui,Xia Changsheng,Wang Ziwei,Zhu Yanxi,Wang Yifan,Chaihu Lingxiao,Tan Minjie,Wang Hongwei,Huang Yanyi,Wang Jianbin,Wang Guanbo

The characterization of protein complex is vital for unraveling biological mechanisms in various life processes. Despite advancements in biophysical tools, the capture of non-covalent complexes and deciphering of their biochemical composition continue to present challenges for low-input samples. Here we introduce SNAP-MS, a Stationary-phase-dissolvable Native Affinity Purification and Mass Spectrometric characterization strategy. It allows for highly efficient purification and characterization from inputs at the pico-mole level. SNAP-MS replaces traditional elution with matrix dissolving during the recovery of captured targets, enabling the use of high-affinity bait-target pairs and eliminates interstitial voids. The purified intact protein complexes are compatible with native MS, which provides structural information including stoichiometry, topology, and distribution of proteoforms, size variants and interaction states. An algorithm utilizes the bait as a charge remover and mass corrector significantly enhances the accuracy of analyzing heterogeneously glycosylated complexes. With a sample-to-data time as brief as 2 hours, SNAP-MS demonstrates considerable versatility in characterizing native complexes from biological samples, including blood samples.