Poster Presentation 27th Annual Lorne Proteomics Symposium 2022

Multiplexing dimethyl labelled samples to enhance MS1 quantitation in single-cell proteomics (#113)

Toby A Dite 1 , Samantha J Emery-Corbin 1 , Laura F Dagley 1 , Andrew Webb 1
  1. Walter and Eliza Hall Institute, Parkville, VIC, Australia

The frontier of single-cell proteomics is being defined by new methods of LC-MS that allow for quantification of low-abundant samples. By employing a combination of miniaturized sample preparation, nano-flow chromatography and DIA-PASEF, thousands of proteins can be identified from sub-nanogram amounts of starting material (1). In addition, sample losses can be minimized and MS2 sequencing in DDA can be augmented by multiplexing low-abundant samples with ‘carrier’ peptides using isobaric labelling (2,3). Similarly, non-isobaric (isotopologous) tags such as dimethyl labelling and mTRAQ can allow for sample multiplexing (4). These methods necessitate DIA as the numbers of distinguishable precursors are increased in the multiplexed sample, making DDA less amenable for sample throughput. It has been shown previously that high sample throughput can be achieved using mTRAQ labelling and DIA (5). Here, we aimed to determine whether DIA-PASEF could be used as a method for increasing proteome coverage by multiplexing samples with light, medium and heavy dimethyl labels, and ultimately improve quantitative information for use in single cell proteomics.

 

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