RePlay^® for nanoLC/MS

Identify 20% to 40% more proteins than nanoLC/MS alone. RePlay provides proteomic researchers the ability to analyze a single nanoLC injection twice without the need for additional time or sample. Obtain maximum information from a single nanoLC/MS run for more coverage and increased analytical confidence.

RePlay uses a nanoflow splitter and the unique RePlay valve to split the nanoLC eluent to a direct on-line analysis and to a Capture Cartridge. During equilibration of the column, the RePlay valve switches to direct the captured chromatogram to the MS for a second analysis of the same injection.

• Use Existing Equipment - RePlay is compatible with existing MS, nanoLC, and nanoelectrospray ionization sources (e.g., nanospray needles, pulled capillaries, ESI Chip, etc.).
  
  
• Twice the Possibilities - RePlay performs a second analysis of one sample without a reinjection, thereby allowing for twice the analytical possibilities from every nanoLC injection. Use the second analysis to obtain more targeted information.
  
  
• No Additional Time - RePlay splits the sample from one injection, so a second analysis is provided with no additional timerequired.
  
  
• No Additional Sample - Utilizing the concentration-sensitive nature of electrospray, RePlay can split the sample to two slower flows at the same concentration without decrease in overall signal intensity.

RePlay analysis enables the following analytical possibilities:

• Orthogonal MS Experiments - CID, ETD, HCD, IRMPD, ECD, MS3, precursor ion, etc.
  
  
• Separate Mass Ranges - Separate the mass ranges into the direct and RePlay analyses to identify more ions within each mass range.
  
  
• Improved Resolution - Slow down the chromatography during the RePlay analysis to improve chromatographic resolution with the use of a refocusing column.
  
  
• Vary MS Conditions - Use a different collision energy during the RePlay analysis to improve MS/MS sensitivity.
  
  
• Focused Analysis - Identify peaks of interest in the first analysis and use the RePlay analysis to dig deeper.
  
  
• Exclusion and Inclusion List - Use exclusion and inclusion lists to look at additional ions or target those without a positive identification during the RePlay analysis.
  
  
• Different Charge States - Look at doubly charged ions in the first analysis and triply charged and greater ions during the RePlay analysis to enhance ion fragmentation and improve structural information.
  
  
• Extended Analysis - Slow down the chromatography without a refocusing column to broaden peak widths and provide more time during the RePlay analysis for data mining.
  
  
• Positive and Negative Ion Analysis - Analyze in negative ion mode to identify PTMS, then switch to positive ion mode during the RePlay analysis to increase structural information.
  
  
• Exploit Random Gains - Improve coverage from the random gains that generally occur during a repeat analysis of the same sample.
  
  
• Reproducibility - Increase analytical completeness with a duplicate run for higher confidence.

Request copy of poster: Combining nanoLC-RePlay®-MS/MS with a Scheduled Precursor List (SPL, Exclusion/Inclusion List) to Significantly Enhance Protein Identification From Complex Biological Samples

Authors: 1. Daniel Eikel; 2. Christian Albers and Markus Lubeck 1. Advion BioSystems, Inc., Ithaca, NY, USA; 2. Bruker Daltonik GmbH, Bremen, Germany