Selected reaction monitoring assays
Provided herein are methods for developing selected reaction monitoring mass spectrometry (LC-SRM-MS) assays.
1. A composition comprising isolated synthetic peptides said composition comprising ALQASALK (SEQ ID NO: 58), GFLLLASLR (SEQ ID NO: 1644), LGGPEAGLGEYLFER (SEQ ID NO: 596), VEIFYR (SEQ ID NO: 941), INPASLDK (SEQ ID NO: 153) and TSYQVYSK (SEQ ID NO: 155).
2. A composition comprising at least 10 isolated synthetic peptides said composition comprising at least 2 isolated synthetic peptides of each TSP1, ALDOA, FRIL, LG3BP, and C163A, and wherein the at least two isolated synthetic peptides are selected from ADDGRPFPQVIK (SEQ ID NO: 57), ALQASALK (SEQ ID NO: 58), QLLLTADDR (SEQ ID NO: 60), ALFQDIK (SEQ ID NO: 593), DDV ALEGVSHFFR (SEQ ID NO: 594), KPAEDEWGK (SEQ ID NO: 595), LGGPEAGLGEYLFER(SEQ IDNO: 596), LNQALLDLHALGSAR (SEQ ID NO: 597), MGDHLTNLHR (SEQ ID NO: 598), ASHEEVEGL VEK (SEQ ID NO: 938), STHTLDLSR (SEQ ID NO: 940), VEIFYR (SEQ ID NO: 941), YSSDYFQAPSDYR (SEQ ID NO: 942), FQDLVDA VR (SEQ ID NO: 1641), FTGSQPFGQGVEHATANK (SEQ ID NO: 1642), FVFGTIPEDILR (SEQ ID NO: 1643), GFLLLASLR (SEQ ID NO: 1644), INPASLDK (SEQ ID NO: 153), LEVFYNGAWGTVGK (SEQ ID NO: 154), TSYQVYSK (SEQ ID NO: 155), and GTSQNDPNWVVR (SEQ ID NO: 1645).
3. The composition of claim 2 , wherein at least one of the isolated synthetic peptides is isotopically labeled.
4. The composition of claim 2 , wherein the measured quantity of each of the at least 2 isolated synthetic peptides of each TSP1, ALDOA, FRIL, LG3BP, and C163A is known.
5. The composition of claim 2 , further comprising one or more polar solvents.
6. The composition of claim 2 , further comprising an additional five isolated synthetic peptides of at least one protein selected from LRP1, BGH3, COIA1, TETN, TSP1, ALDOA, GRP78, ISLR, FRIL, LG3BP, PRDX1, FIBA, KIT, GGH, C163A and GSLGI.
7. The composition of claim 2 , further comprising an additional five isolated synthetic peptides of at least one protein selected from APOE, BASP1, CD14, FOXA2 or HSPB 1.
8. A composition comprising at least 25 isolated synthetic peptides said composition comprising at least five isolated synthetic peptides of each TSP1, ALDOA, FRIL, LG3BP, and C163A wherein the at least five isolated synthetic peptides are selected from ADDGRPFPQVIK (SEQ ID NO: 57), ALQASALK (SEQ ID NO: 58), QLLLTADDR (SEQ ID NO: 60), ALFQDIK (SEQ ID NO: 593), DDV ALEGVSHFFR (SEQ ID NO: 594), KPAEDEWGK (SEQ IDNO: 595), LGGPEAGLGEYLFER(SEQ IDNO: 596), LNQALLDLHALGSAR (SEQ ID NO: 597), MGDHLTNLHR (SEQ ID NO: 598), ASHEEVEGLVEK (SEQ ID NO: 938), STHTLDLSR (SEQ ID NO: 940), VEIFYR (SEQ ID NO: 941), YSSDYFQAPSDYR (SEQ ID NO: 942), FQDLVDAVR (SEQ ID NO: 1641), FTGSQPFGQGVEHATANK (SEQ ID NO: 1642), FVFGTIPEDILR (SEQ ID NO: 1643), GFLLLASLR (SEQ ID NO: 1644), INPASLDK (SEQ ID NO: 153), LEVFYNGAWGTVGK (SEQ ID NO: 154), TSYQVYSK (SEQ ID NO: 155), and GTSQNDPNWVVR (SEQ ID NO: 1645).
9. The composition of claim 8 , wherein the at least five isolated synthetic peptides are selected from GFLLLASLR (SEQ ID NO: 1644), LGGPEAGLGEYLFER (SEQ ID NO: 596), ALQASALK (SEQ ID NO: 58), VEIFYR (SEQ ID NO: 941), INPASLDK (SEQ ID NO: 153), LEVFYNGAWGTVGK (SEQ ID NO: 154), and TSYQVYSK (SEQ ID NO: 155).
10. A method comprising analyzing the composition of claim 8 using mass spectrometry.
11. The method of claim 10 , wherein the method uses selected reaction monitoring mass spectrometry.
12. A method comprising analyzing the composition of claim 8 using Enzyme-Linked Immunosorbent Assay (ELISA).