BCL-2-like protein 11 SRM/MRM assay
First Claim
1. A method for measuring the level of the human Bcl-2-like protein 11 (BIM) in a human biological sample of formalin-fixed tissue, comprising detecting and quantifying the amount of a BIM fragment peptide in a protein digest prepared from said biological sample using mass spectrometry;
- and calculating the amount of BIM protein in said sample;
wherein the BIM fragment peptide consists of the peptide of SEQ ID NO;
2 andwherein said level is a relative level or an absolute level.
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Abstract
Specific peptides, and derived ionization characteristics of those peptides, from the Bcl-2-like protein 11 (BIM) are provided that are particularly advantageous for quantifying the BIM protein directly in biological samples that have been fixed in formalin by the method of Selected Reaction Monitoring (SRM) mass spectrometry, or what can also be termed as Multiple Reaction Monitoring (MRM). Such biological samples are chemically preserved and fixed where the biological sample is selected from tissues and cells treated with formaldehyde containing agents/fixatives including formalin-fixed tissue/cells, formalin-fixed/paraffin embedded (FFPE) tissue/cells, FFPE tissue blocks and cells from those blocks, and tissue culture cells that have been formalin fixed and or paraffin embedded. A protein sample is prepared from the biological sample using the Liquid Tissue™ reagents and protocol, and the BIM protein is quantitated in the Liquid Tissue™ sample by the method of SRM/MRM mass spectrometry by quantitating in the protein sample at least one or more of the peptides described. These peptides can be quantitated if they reside in a modified or an unmodified form. An example of a modified form of a BIM peptide is phosphorylation of a tyrosine, threonine, serine, and/or other amino acid residues within the peptide sequence.
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Citations
12 Claims
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1. A method for measuring the level of the human Bcl-2-like protein 11 (BIM) in a human biological sample of formalin-fixed tissue, comprising detecting and quantifying the amount of a BIM fragment peptide in a protein digest prepared from said biological sample using mass spectrometry;
- and calculating the amount of BIM protein in said sample;
wherein the BIM fragment peptide consists of the peptide of SEQ ID NO;
2 andwherein said level is a relative level or an absolute level. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
- and calculating the amount of BIM protein in said sample;
Specification