Isobaric Mass Tags for Proteomic Multiplexing
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Solution Overview
Problem
Current isobaric mass tags have limitations in the range of molecules they can label and the levels of multiplex analysis achievable, leading to increased complexity in mass spectra and reduced sensitivity in proteomic studies.
Innovation Solution
Development of novel isobaric mass tags with a common core structure based on DMPip-βAla, allowing selective labelling properties and additional offset masses, which can convert to isochemic tags for quantitation in LC-MS and direct isobaric mass tags for biomarker qualification, using reactive functionalities like sulfo-tetrafluorophenyl and beta-alanine moieties to enhance multiplexing rates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If isobaric mass tags are used to reduce mass spectrum complexity, then measurement precision is improved, but device complexity increases due to requiring MS/MS fragmentation and multiple sample sets
Solution Approach 1:
The mass tag is divided into distinct functional segments: a common core structure (DMPip-βAla) that provides the mass normalization and a variable mass series modifying group that provides sample-specific offset masses. This segmentation allows the common core to be reused across multiple tag sets while differentiating samples through the variable groups, reducing the need for entirely different tag chemistries for each sample set.
Solution Approach 2:
The common core structure serves multiple functions simultaneously: it provides mass normalization across all samples, acts as the basis for isobaric tagging (same aggregate mass), and can be converted to isochemic tags for LC-MS quantitation. This multi-functionality reduces the number of different tag chemistries needed and simplifies the overall workflow.
2Productivity
If multiple isobaric mass tag sets are developed to increase multiplexing rates, then productivity is improved, but device complexity and manufacturing cost increase
Solution Approach 1:
A single common core structure serves as the foundation for multiple isobaric mass tag sets by combining with different mass series modifying groups. This universal core approach allows generation of multiple tag sets (e.g., 6-plex, 12-plex, 18-plex) without developing entirely new chemistries for each set, thereby increasing multiplexing capacity while controlling complexity and cost.
Solution Approach 2:
Multiple mass series modifying groups are combined with the common core structure to create different tag sets with different offset masses. This merging strategy allows multiple samples to be analyzed simultaneously using a unified tag platform, increasing productivity while avoiding the need for separate tag development for each sample set.
3Ease of operation
If existing amine labelling reagents are used, then ease of operation is maintained, but adaptability is limited to amine-containing molecules
Solution Approach 1:
Different reactive functionalities are introduced at specific positions in the mass tag structure to target different molecular functional groups. For example, hydrazide groups can react with carbonyls in steroids, while the common core structure remains unchanged. This allows the same platform to label diverse molecule types (proteins, steroids, nucleic acids) by simply changing the reactive group, thereby increasing adaptability while maintaining operational simplicity.
Data Source
AI summary
A reactive mass label for labelling a biological molecule for detection by mass spectrometry, which label comprises a reactive functionality for labelling thiol groups or carbonyl groups. Also provided is a reactive mass label for labelling a biological molecule for detection by mass spectrometry, wherein the mass label comprises the following structure:X-L-M-S—Rewherein X is a mass marker moiety, L is a cleavable linker, M is a mass normalization moiety, S is a mass series modifying group comprising the following group:wherein J is C═O, K is NH, and n is 2 or J and K are both CH2 and n is 1, and wherein m is at least 1; and Re is a reactive functionality for attaching the mass label to a biological molecule.