High Mass Linear Analysis of Intact Proteins on the 4800...
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www.appliedbiosystems.com Technical Note High Mass Linear Analysis of Intact Proteins on the 4800 MALDI TOF/TOF™ Analyzer Introduction The mass measurement of intact proteins is an important first step in protein quality control, identifying proteins and identification of biomolecular interactions. This capability is enabled by the high sensitivity linear detection system on the 4800 MALDI TOF/TOF™ Analyzer, complimenting its exceptional low mass MS and MS/MS performance. Key Benefits •High sensitivity in-line linear detector provides unparalleled high mass data •Equivalent linear mode performance to the industry standard Voyager™ series of instruments •Compliments the outstanding low mass performance of the 4800 MALDI TOF/TOF™ Analyzer Discussion The high mass linear mode performance linear mode was demonstrated with Bovine Serum Albumin (BSA) (66,431 m/z), Immunoglobulin 1 (IgG1) (149,500 m/z), and a mixture containing BSA, IgG1, and Aldolase (39,105 m/z) using the default high mass linear methods on the 4800 MALDI TOF/TOF™ Analyzer. For comparative purposes the sample were also analyzed on a Voyager-DE PRO instrument. 15000 42000 69000 96000 123000 150000 Mass (m/z) 3565.5 0 20 40 60 80 100 % Intensity 15000 42000 69000 96000 123000 150000 Mass (m/z) 3565.5 0 20 40 60 80 100 % Intensity 15000 42000 69000 96000 123000 150000 Mass (m/z) 1.4E+4 0 20 40 60 80 100 % Intensity 15000 42000 69000 96000 123000 150000 Mass (m/z) 1.4E+4 0 20 40 60 80 100 % Intensity 33216 66431 dimer dimer 33216 66431 Voyager DE Pro 4800 MALDI TOF/TOF™ Analyzer Figure 1 High Mass Linear Mode analysis of BSA in sinapinic acid. Singly and doubly charged BSA is observed at m/z 66,431 and m/z 33,216 respectively. A BSA dimer is also observed. The top panel represents data collected on a Voyager DE Pro. The bottom panel represents data collected on a 4800 MALDI TOF/TOF™ Analyzer.
Transcript of High Mass Linear Analysis of Intact Proteins on the 4800...
www.appliedbiosystems.com
Technical Note
High Mass Linear Analysis of Intact Proteins on the 4800 MALDI TOF/TOF™Analyzer
Introduction
The mass measurement of intact proteins is an important first step in protein quality control, identifying proteins and identification of biomolecular interactions. This capability is enabled by the high sensitivity linear detection system on the 4800 MALDI TOF/TOF™ Analyzer, complimenting its exceptional low mass MS and MS/MS performance.
Key Benefits•High sensitivity in-line linear detector provides unparalleled high mass data
•Equivalent linear mode performance to the industry standard Voyager™ series of instruments
•Compliments the outstanding low mass performance of the 4800 MALDI TOF/TOF™Analyzer
DiscussionThe high mass linear mode performance linear mode was demonstrated with Bovine Serum Albumin (BSA) (66,431 m/z), Immunoglobulin 1 (IgG1) (149,500 m/z), and a mixture containing BSA, IgG1, and Aldolase (39,105 m/z) using the default high mass linear methods on the 4800 MALDI TOF/TOF™ Analyzer. For comparative purposes the sample were also analyzed on a Voyager-DEPRO instrument.
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Figure 1 High Mass Linear Mode analysis of BSA in sinapinic acid. Singly and doubly charged BSA is observed at m/z 66,431 and m/z 33,216 respectively. A BSA dimer is also observed. The top panel represents data collected on a Voyager DE Pro. The bottom panel represents data collected on a 4800 MALDI TOF/TOF™ Analyzer.
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Technical Note
Example 1, BSA (4 pmol/uL), was prepared and deposited onto a MALDI plate using sinapinic acid as matrix. Three peaks are identified in the spectra. The peak at m/z 66,431 represents singly charged BSA, the peak at m/z 33,216 represents doubly charged BSA, and the small peak at ≈133,000 m/z represents a BSA dimer. Figure 1 is the result of an experiment performed on the deposited BSA comparing high mass linear mode analysis on a Voyager DE PRO with similar analysis on a 4800 MALDI TOF/TOF™ Analyzer. The top panel of Figure 1 shows data collected on a Voyager-DE PRO. The bottom panel of Figure 1 shows data collected on a 4800 MALDI TOF/TOF™ Analyzer. The high mass linear default methods (focus mass for IgG) were used to collect this data.
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Figure 2 High Mass Linear Mode analysis of IgG1 in sinapinic acid. Singly and doubly charged IgG1 is observed at m/z 178,500 and m/z 74,249 respectively. The top panel represents data collected on a Voyager-DE Pro. The bottom panel represents data collected on a 4800 MALDI TOF/TOF™ Analyzer.
Example 2, IgG1 (0.6 pmol/uL), was prepared and deposited onto a MALDI plate using sinapinicacid as matrix. Two peaks are identified in the spectra. The peak at m/z 148,500 represents singly charged IgG1 and the peak at m/z 74,249 represents doubly charged IgG1. Figure 2 is the result of an experiment performed on the deposited IgG1 comparing high mass linear mode analysis on a Voyager-DE PRO with similar analysis on a 4800 MALDI TOF/TOF™ Analyzer. The top panel of Figure 2 shows data collected on a Voyager-DE PRO. The bottom panel of Figure 2 shows data collected on a 4800 MALDI TOF/TOF™ Analyzer. The high mass linear default (factory) method (focus mass for IgG) was used to collect this data.
Voyager DEPro
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Technical Note
Figure 3 High Mass Linear Mode analysis of a High Mass Linear mixture of intact proteins in sinapinic acid. The top panel represents data collected on a Voyager DE™ Pro. The bottom panel represents data collected on a 4800 TOF/TOF™ Analyzer.
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Example 3, High Mass Linear mixture (HML) (rabbit aldolase (176 fmol/uL), BSA (200 fmol/uL), and IgG 1(600 fmol/uL)) was prepared and deposited onto a MALDI plate using sinapinic acid as matrix. Multiple peaks are identified in the spectra. The peak at m/z 39,105 represents aldolase, the peaks at m/z 66,431 and 33,214 represent singly and doubly charged BSA. The peaks at m/z 148,500 and 74,249 represent singly and doubly charged IgG1. Figure 3 is the result of an experiment performed on the deposited HML mixture analyzed using the default high mass linear methods on a Voyager-DE PRO and a 4800 MALDI TOF/TOF™ Analyzer. The top panel of Figure 3 shows data collected on a Voyager-DE PRO. The high mass linear default (factory) method (IgG) was used to collect this data. The bottom panel of Figure 3 shows data collected on a 4800 MALDI TOF/TOF™ Analyzer. The high mass linear default method (focus mass for IgG) was used to collect this data.
High quality, high mass linear analysis is demonstrated on the 4800 MALDI TOF/TOF™Analyzer. This has been shown using intact proteins BSA and IgG1, and a High Mass Linear mixture containing aldolase, BSA, and IgG1. Similar data quality is achieved to that on a Voyager-DE PRO. The 4800 MALDI TOF/TOF™ has the added benefits of high resolution and TOF/TOF™ capability allowing laboratories the flexibility to required for solving all their biological problems.
Summary
Applera Corporation is committed to providing the world’s leading technology and information for life scientists. Applera Corporation consists of the Applied Biosystems and Celera Genomics businesses. MALDI TOF/TOF, Voyager, Voyager-DE and TOF/TOF are trademarks of Applied Biosystems/MDS SCIEX, a joint venture between Applera Corporation and MDS Inc. AB (Design), Opti-TOF, iScience, iScience (Design) and Applera are trademarks, and Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the U.S. and/or certain other countries. MDS and SCIEX are registered trademarks of MDS Inc. All other trademarks are the property of their respective owners.For Research Use Only. Not for use in diagnostic procedures. 2005 Applera Corporation and MDS Inc. All rights reserved. Information subject to change without notice.
