Methods and compositions for mass spectrometry analysis
a mass spectrometry and composition technology, applied in the field of methods and compositions for mass spectrometry analysis and mass spectrometry, can solve the problems of loss of analytes, no prior art patents used modified matrix,
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example 1
[0061]A DHB like molecule (photon absorbing moiety) is coupled with a lipophilic long alkyl chain (affinity moiety), therefore has affinity for lipophilic compounds (FIG. 1). This affinity matrix could selectively desorb / ionize lipophilic analyte in a mixture for MALDI mass analysis. Using this affinity matrix as matrix and standard MALDI analysis protocol (protocol available from Mass Spectrometry for Biotechnology; Gary Siuzdak, Academic Press 1996), a sample containing a mixture of dynorphin A-(1-11) and more lipophilic acetylated dynorphin A-(1-11) at 1:1 ratio gave 10 times higher peak of acetylated dynorphin A-(1-11) than the peak of less lipophilic dynorphin A-(1-11) while using DHB as matrix gave almost same peak height for two analytes. This enhanced signal of acetylated dynorphin A-(1-11) indicates the selective desorption / ionization capability of the lipophilic affinity matrix. The typical mixing and incubation time is several minutes. Longer incubation time can result in...
example 2
[0062]FIG. 2 shows a polymer having both affinity groups and photon energy absorbing groups covalently coupled with it. The polymer 1 is polylysine (MW=20,000), the photon energy absorbing groups 2 are alpha-cyano-4-hydroxycinnamic acid (CCA) molecules and the affinity groups 3 are antibodies. The CCA and antibodies are coupled to the side chains of polylysine via amide bonds. The preferred ratio of antibody to CCA is 1:5 to 1:20. This polymer can be used as a selective affinity matrix to selectively desorb / ionize the corresponding antigen in MALDI analysis. A further modification of this affinity matrix is that the affinity groups are covalently linked to the polymer back bone while the photon energy absorbing groups are bounded to the polymer by non-covalent interaction such as ion pairing or lipophilic interaction.
example 3
[0063]FIG. 3 shows the examples of several reactive matrix molecules: 2,5-dihydroxybenzoic acid (DHB)-NHS ester, alpha-cyano-4-hydroxycinnamic acid (CCA)-NHS ester and 3-picolinic acid-NHS ester. The DHB-NHS ester 4, 3-Picolinic acid-NHS ester 5 and CCA-NHS ester 6 are active esters of known matrix DHB, CCA and 3-Picolinic acid respectively. They can react with the analyte molecules containing free amine groups upon mixing and incubation in sample solution. Preferred incubation time is 10˜60 minutes. Using these reactive matrix molecules as matrix and standard MALDI analysis protocol, the analyte containing amine groups can be readily detected in MALDI analysis. Reactive matrix can also be immobilized on solid phase support such as the structure 7 in the figure, in structure 7, the 4, 3-picolinic acid-NHS ester is immobilized on a PEG resin (Nova biochem), therefore allow easy purification of unreacted matrix.
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