Combining ultra-high resolution ion mobility spectrometry with cryogenic IR spectroscopy for the study of biomolecular ions

Abstract

We explore the capability of SLIM-based IMS for isomer selectivity in combination with cryogenic, messenger-tagging IR spectroscopy.

Analyzing glycans cleaved from a biotherapeutic protein using ultrahigh-resolution ion mobility spectrometry together with cryogenic ion spectroscopy

“…Ion-mobility-selective IR spectroscopy Analysis of N-linked glycan standards as well as those enzymatically cleaved from etanercept was performed on a homebuilt instrument that combines ultrahigh-resolution ion mobility spectrometry and cryogenic vibrational spectroscopy, as described in a series of recent publications. 23,25,26,36 Briefly, doubly protonated glycan ions are generated by a nanoelectrospray ionization (nESI) source and introduced into the first vacuum stage through a heated stainless steel capillary. After exiting the capillary, the ions enter a dual ion funnel trap (IFT), where they are focused and accumulated into packets.…”

“…6,[10][11][12][13][14][15][16][17] Recently, spectroscopy-based methods have been combined with MS to tackle the issue of glycan identification. [18][19][20][21][22][23][24][25][26][27][28] For relatively large and complex molecules such as N-linked glycans, cryogenic spectroscopy is best suited to distinguishing the subtle structural differences between isomers. [21][22][23][24][25][26][27][28][29] Because vibrational spectra reflect intrinsic molecular properties of a molecule, they are insensitive to the range of experimental conditions that might be encountered in different laboratories.…”

“…[18][19][20][21][22][23][24][25][26][27][28] For relatively large and complex molecules such as N-linked glycans, cryogenic spectroscopy is best suited to distinguishing the subtle structural differences between isomers. [21][22][23][24][25][26][27][28][29] Because vibrational spectra reflect intrinsic molecular properties of a molecule, they are insensitive to the range of experimental conditions that might be encountered in different laboratories. 26 In this work, we combine ultrahigh-resolution ion mobility spectrometry and cryogenic, vibrational spectroscopy to analyze four of the N-glycans released from the therapeutic protein etanercept, which is a key pharmaceutical component of the drug Enbrel™.…”

“…[21][22][23][24][25][26][27][28][29] Because vibrational spectra reflect intrinsic molecular properties of a molecule, they are insensitive to the range of experimental conditions that might be encountered in different laboratories. 26 In this work, we combine ultrahigh-resolution ion mobility spectrometry and cryogenic, vibrational spectroscopy to analyze four of the N-glycans released from the therapeutic protein etanercept, which is a key pharmaceutical component of the drug Enbrel™. Etanercept is a dimeric fusion protein containing the extracellular domain of the human tumor necrosis factor receptor (TNFR) attached to the fragment crys-tallizable (Fc) portion of human immunoglobulin heavy chain G1 (IgG1) 30 by disulfide bonds (and hence its abbreviation TNFR-Fc).…”

How General Is Anomeric Retention during Collision-Induced Dissociation of Glycans?

“…3(a, b)), which we separately measured after separation by SLIM-based ion mobility. 33 The spectra of the C2 fragments from both LNnT (Fig. 3(c)) and LNnH ( Fig.…”