MSP BigAnchor Targets BC

Transcription

1 Instructions for Use MSP BigAnchor Targets BC Polished steel targets with hydrophilic sample and calibrant spots surrounded by a hydrophobic barrier for MALDI sample preparation CARE products are designed to support our worldwide customers with highquality consumables, accessories and dedicated kits. The CARE product range is specifically optimized and certified for use with all Bruker Daltonics systems. Language: en Revision C (December 2015)

2 1 Product Description 2 2 MSP BigAnchor Targets BC Product Overview 3 3 Software Requirements 4 4 Risk and Safety Information 4 5 General Sample Preparation Recommendations for MSP BigAnchor Targets BC Compatibility of Solvents with the Hydrophobic Coating of MSP BigAnchor Targets BC 5 6 Sample Preparation Protocols for BigAnchor Targets (Ø = 2000 µm) HCCA (Dried Droplet) Preparation for Peptides and Protein Digests ,5-DHB (Dried Droplet) Preparation for Peptides, Phospho- and Glycoprotein Digests, Intact Proteins, and Glycans Sinapinic Acid (Double Layer) Preparation for Intact Proteins ,5-Diaminonaphthalene (DAN) Preparation for Top-down Sequencing of Proteins 8 7 Cleaning MSP BigAnchor Targets BC 9 8 Instrument Teaching of Anchor Positions 9 9 Calibrant Anchors 9 10 Manufacturer 10 1 Product Description Sample positions on patented 1 AnchorChip targets contain "anchors"; hydrophilic patches surrounded by a hydrophobic ring. The "anchor" localizes droplets at the sample position and the hydrophobic ring prevents sample spreading and concentrates the sample into a spot between 0.4 and 2 mm in diameter. After correct adjustment of the target in the MALDI ion source, the localization effect ensures that every single laser shot fired throughout an automatic run will hit a sample spot. This significantly increases the efficiency of the MALDI acquisition process. In addition, the concentration effect provides enhanced sensitivity when analyzing dilute samples (see Figure 1). The hydrophobic coating not only repels aqueous liquids, but also polar organic solvents (for example, alcohols, acetonitrile and acetone). Therefore, MSP BigAnchor Targets BC are well-suited for use with sample solutions with a high organic solvent content, which would otherwise spread over the target. It is important that the protocols in these Instructions for Use are followed rigorously. Deviating from the given procedures may adversely affect the hydrophobic coating and reduce the number of times that the target can be reused. 1 Bruker s AnchorChip Technology is protected by German, UK and US patents. Patents applied for elsewhere. Page 2 of 10 MSP BigAnchor Targets BC Instructions for Use Revision C

3 Figure 1 The basic principle of MSP BigAnchor Targets BC 2 MSP BigAnchor Targets BC Product Overview MSP BigAnchor Targets BC are used in the microscout ion source that is found in microflex series instruments. They can also be used in autoflex and ultraflex series instruments when mounted in the MTP MSP adapter (# ). MSP BigAnchor Targets BC with barcode for microflex instruments Part No Product Name Spot Ø (µm) Sample Spots Calibrant Spots MSP BigAnchor 24 BC / MSP BigAnchor 96 BC The barcode on the target consists of a ten-digit number: the first three digits indicate the target type and the last seven digits constitute a unique serial number. The barcode can be read using the Honeywell 3800G CCD Handscanner (Bruker # ) 1. Laser barcode scanners cannot be used to read the barcode on Bruker MALDI targets. 1 Other CCD barcode scanners may be able to read the barcode, but Bruker does not guarantee that all CCD scanners are compatible with Bruker MALDI targets. Revision C MSP BigAnchor Targets BC Instructions for Use Page 3 of 10

4 3 Software Requirements Compass for flex series instruments software (Version 1.3 and later) provides full support (geometry, calibration templates) for all MSP BigAnchor targets. Earlier versions require a software patch or update to support BigAnchor targets. Contact MALDI_SW_Support@bruker.com for more information. 4 Risk and Safety Information Additional chemicals may be required for procedures described in these Instructions for Use. Carefully read the Material Safety Data Sheet provided by the supplier and follow general safety regulations when handling chemicals or biohazardous material. 5 General Sample Preparation Recommendations for MSP BigAnchor Targets BC Consumables Many plastics are not compatible with mass spectrometric analysis because they release polymers. Avoid all types of siliconized ( low- bind ) tubes. Using unsuitable plasticware typically results in heterogeneous crystallization and polymer signals, which can adversely affect analyses. In extreme cases, samples do not crystallize at all and remain liquid even after many hours. Table 1 Recommended consumables for MSP BigAnchor Targets BC applications Consumables Pipette tips Microcentrifuge tubes Plastic bottles Microplates (MTPs) Recommended supplier Eppendorf tips μl (# , Eppendorf), μl (# , Eppendorf), μl (# , Eppendorf) Eppendorf tubes, 1.5 ml (# , Eppendorf) Nalgene FEP Teflon bottles Greiner PP, natural MTP Solvents and Chemicals Solvents and chemicals used for MALDI-TOF-MS must be high-purity reagents (at least HPLC-grade). Avoid the use of detergents. Page 4 of 10 MSP BigAnchor Targets BC Instructions for Use Revision C

5 5.1 Compatibility of Solvents with the Hydrophobic Coating of MSP BigAnchor Targets BC The hydrophobic coating of MSP BigAnchor Targets BC is resistant to water and polar organic solvents (for example, alcohols, acetonitrile, and acetone) at neutral and acidic ph. CAUTION Table 2 Do not use solvents or solutions with a basic ph. Using solutions with a basic ph will destroy the hydrophobic coating of MSP BigAnchor Targets BC. Solvents and chemicals compatible with MSP BigAnchor Targets BC Solvents 2-propanol Water Acetonitrile Ethanol Other chemicals Trifluoroacetic acid (TFA) Ammonium dihydrogen phosphate Sodium chloride Diammonium hydrogen citrate If your workflow requires a solvent or chemical not listed in Table 2, we strongly recommend that the chemical compatibility is checked. Deposit a 1 µl droplet outside the sample spot region of the target and incubate for 5 minutes. After 5 minutes, aspirate the liquid and check that the coating is still hydrophobic at the test location. Revision C MSP BigAnchor Targets BC Instructions for Use Page 5 of 10

6 6 Sample Preparation Protocols for BigAnchor Targets (Ø = 2000 µm) Sample preparation protocols for BigAnchor targets are similar to conventional dried droplet protocols used for steel targets. The protocols differ in the amount of matrix added to each target position: BigAnchor protocols require only a quarter of the amount of matrix used in conventional protocols. Note The following sample preparation procedures should not be used if your workflow contains a prescribed sample preparation method (for example, IVD MALDI Biotyper SOP). The following solvents and chemicals are recommended for BigAnchor target sample preparation procedures. Solvents Ultrapure water, CHROMASOLV for HPLC (Sigma-Aldrich # ) Acetonitrile 99.9%, LC-MS CHROMASOLV grade at Fluka (Sigma-Aldrich # 34967) Ethanol absolute, 99.8% (GC), (undenaturated) (Sigma-Aldrich # 24102) Other chemicals Trifluoroacetic acid >98,5% ( ReagentPlus grade) (Sigma-Aldrich # T62200) Sodium chloride; NaCl (required for glycan analysis, see section 6.2) 6.1 HCCA (Dried Droplet) Preparation for Peptides and Protein Digests This protocol uses α-cyano-4-hydroxycinnamic acid (HCCA # or # ) as a MALDI matrix. Chemicals and materials required Matrix stock solution a saturated solution of HCCA in TA30 (30:70 [v/v] acetonitrile: 0.1% TFA) Matrix working solution matrix stock solution diluted 1:4 in TA30 Sample solvent 0.1% TFA or TA30 Sample preparation protocol 1. Mix 1 part sample solution with 1 part matrix working solution. 2. Deposit 1 µl of the sample/matrix mixture onto the BigAnchor target and allow to dry. Page 6 of 10 MSP BigAnchor Targets BC Instructions for Use Revision C

7 6.2 2,5-DHB (Dried Droplet) Preparation for Peptides, Phospho- and Glycoprotein Digests, Intact Proteins, and Glycans This protocol uses 2,5-Dihydroxybenzoic acid (2,5-DHB; # ) as a MALDI matrix. Chemicals and materials required Sample solvent 0.1% TFA or TA30 (30:70 [v/v] acetonitrile: TFA 0.1% in water) o For glycan analysis, use water as the sample solvent Matrix solution 30 g/l 2,5-DHB in TA30 o For glycan analysis, supplement the matrix solution with 1 mm NaCl Sample preparation protocol 1. Mix 1 part sample solution with 1 part matrix solution. 2. Deposit 2 µl of the sample/matrix mixture onto the BigAnchor target and allow to dry. 6.3 Sinapinic Acid (Double Layer) Preparation for Intact Proteins This protocol uses sinapinic acid (SA; # ) as a MALDI matrix. Chemicals and materials required Matrix solution A saturated solution of SA in ethanol Matrix solution B saturated solution of SA in TA30 (30:70 [v/v] acetonitrile: 0.1% TFA) Sample solvent 0.1% TFA Sample preparation protocol 1. Deposit 0.5 µl of matrix solution A onto the BigAnchor target and allow to dry. 2. Mix 1 part matrix solution B with 1 part analyte solution. 3. Deposit 0.5 µl of the matrix/analyte mixture onto the matrix spot and allow to dry. Revision C MSP BigAnchor Targets BC Instructions for Use Page 7 of 10

8 6.4 1,5-Diaminonaphthalene (DAN) Preparation for Top-down Sequencing of Proteins This protocol uses 1,5- Diaminonaphthalene (Acros Organics product No ), as MALDI matrix. Chemicals and materials required Matrix solution saturated solution of DAN in TA50 (50:50 [v/v] acetonitrile:0.1% TFA in water) Sample solvent 0.1% TFA Sample preparation protocol 1. Deposit 1 2 µl of protein solution onto a spot of the BigAnchor sample plate and allow to dry. 2. Where necessary, wash the sample spot with 10 µl of ice-cold 0.1 %TFA solution. 3. Deposit 1 µl of the matrix solution onto the sample spot and allow to dry. Page 8 of 10 MSP BigAnchor Targets BC Instructions for Use Revision C

9 Bruker 7 Cleaning MSP BigAnchor Targets BC If correctly handled, MSP BigAnchor Targets BC can be reused up to 50 times. For cleaning protocols, see the Bruker Guide to MALDI Sample Preparation (Product Number # ), which is available for download at 8 Instrument Teaching of Anchor Positions Precise spot alignment of the target positions is achieved by using a simple 3-point teaching procedure in the flexcontrol software. For details, refer to the flexcontrol online help. Tip Empty target positions are often almost invisible on the camera video display in the flexcontrol software. Therefore, target positions that will be used as positions for target teaching should be spotted with MALDI matrix to enhance their visibility. 9 Calibrant Anchors MSP BigAnchor Targets BC contain a regular pattern of "calibrant anchors" that can be used for the preparation of external calibrants (see Figure 2). These positions provide optimal external calibration accuracy due to the short distance between sample and calibrant spots. This minimizes mass errors that originate from deviations in plate flatness. The nomenclature used for sample and dedicated calibrant spots is illustrated in Figure 2. Sample spot positions form a raster called Chip #0 (example sample spot positions 0_A1, 0_A2, 0_B1, 0_ B2). Calibrant spots form a raster called Chip #1 (example calibrant spot position 1_A1). When processing prepared MSP BigAnchor Targets BC in the instrument control software (flexcontrol; autoxecute run editor), switching between chip #0 and chip #1 rasters allows the operator to access either the sample or calibrant positions for MALDI analysis. Dedicated calibration templates for the different target types are available in the AutoXecute run editor. Figure 2 Revision C Calibrant anchor layout (96 and 384-spot targets) MSP BigAnchor Targets BC Instructions for Use Page 9 of 10

10 10 Manufacturer Bruker Daltonik GmbH Fahrenheitstraße Bremen Germany Support biotyper.support@bruker.com Phone: +49 (0) Fax: +49 (0) Sales Information care@bruker.com Phone: +49 (0) Web: # , # Descriptions and specifications supersede all previous information and are subject to change without notice. Copyright 2015 Bruker Daltonik GmbH Page 10 of 10 MSP BigAnchor Targets BC Instructions for Use Revision C