A 200 high-pressure N2 cylinder will last approximately 25 Proteograph assays. We recommend changing the cylinder around 25% full to avoid low pressure errors during the assay and possible sample loss.
The Proteograph Product Suite features the following products:
Proteograph Assay Kit
A proprietary panel of five engineered nanoparticles for processing up to 16 samples in one 96-well plate
Buffers and reagents for protein lysis, digestion, and peptide purification
Three quality controls to facilitate the comparison of results across different assays and troubleshooting for a specific assay
PQR Labware Kit
Contains all required labware used with SP100 Automation Instrument with automated peptide quantification and reconstituion methods. Enough labware for 4 runs. Designed to work with the Pierce Fluorometric Peptide Assay (sold separately by Thermo Fisher)
Proteograph Analysis Suite (PAS)
A dedicated software solution for processing, analysis, and visualization of proteomics data sets generated by liquid chromatography-mass spectrometry (LC-MS).
Software includes an integrated search engine for identification and annotation of LC-MS data
We do not install the instrument on a user-provided table. The table that comes with the SP100 is customized specifically for instrument stability and operation. Holes placed in the table are designed for critical access to gas lines and waste containers, while grooves in the table are critical for instrument stability and operational performance.
The Proteograph Assay method can run up to 16 samples per run. If 16 samples are not used, deionized (DI) water should be used in empty sample tubes. Each sample is incubated separately with each of the five nanoparticles, resulting in 80 wells of peptides in a 96-well plate.
Yes, daily and weekly maintenance are a part of normal instrument operation. Maintenance protocols help to ensure optimal operating performance of the SP100 and successful processing of target samples.
Yes, the SP100 Instrument should be powered off when not in use.
The total assay time including sample prep and method setup is 7.5 hours.
Peptide quantification of the samples and controls can serve as an indicator for a successful run.
Yes, back-to-back assays can be run during the same day to process a total of 32 samples (16 samples per assay) in one day. Users would need to be present for approximately 30-45 minutes at the beginning of each run for assay preparation and setup and for approximately 30 minutes at the end of each assay to ensure that the peptide collection plate is properly stored, the instrument deck is cleaned up, and for recommended peptide quantification immediately following assay completion.
No, daily maintenance can be performed at the start of each day, however we would suggest running the MPE flush between assays that are run consecutively in the same day.
Based on recommended starting volumes of 250 µl, there may be approximately 50 µl of sample remaining in the sample tubes. These samples will be left at ambient temperature for the duration of the assay and we do not recommend removing samples in the middle of the runs as lifting the front hood will disrupt normal operation of the assay.
Yes, currently the Proteograph assay is designed to be run with the full five nanoparticle panel. Users can decide which samples are submitted for analysis by mass spectrometry.
Seer recommends using a fluorescence-based quantitation method, such as the Pierce Quantitative Fluorometric Peptide Assay (Thermo Fisher, Cat# 23290). This kit combined with a Seer-specific peptide standard is used in the automated peptide quantification method available on the SP100 platform. Other methods for peptide quantification may be used.
Seer recommends using a trap and elute setup when analyzing samples by LC-MS. While this is not required, we have seen increased longevity and stability of analytical columns and MS signal when using a compatible trap column upstream of the user selected analytical column.
The SP100 should be scheduled for a Preventative Maintenance every 6 months. Contact a Seer representative directly to schedule.
Seer has comprehensively analyzed plasma and serum samples on the SP100. Additional samples types, such as CSF, urine and synovial fluid, have also been tested. Please contact firstname.lastname@example.org for additional information.
250-275 μl per sample is needed.
No. The automated assay has been developed and optimized to be run on the SP100 only.
Any mass spectrometer instrument designed for proteomic analysis can fit seamlessly into the Proteograph workflow. Upon request, Seer can provide recommended methods for LC-MS implementation.
The first step of setting up an analysis depends on preference: start either on the Plates page by selecting Add Plate or on the Data Files page by selecting Link to Plate. Alternatively, you can start on the Projects page and select existing samples.
To reanalyze data analyzed using the Proteograph Analysis Suite, go to the Projects page, select the samples you want to reanalyze, and select Analyze. This option is useful for reanalyzing data using a different analysis method.
The Projects page offer two ways to start an analysis: Analyze and Analyze Controls. The Analyze button analyzes all selected samples in a project, including controls. The Analyze Controls button analyzes controls in the project only. Both buttons open a dialog where you can name the analysis, specify the analysis protocol, and input other general information.
The plate map file is a .csv file that specifies the location of each sample and control in a 96-well plate. The instrument control software (ICS) generates the file and automatically populates each column except MS file name. You must specify the MS data file name in the first column of the file, and then upload it to PAS during analysis setup.
The sample description file is an optional .csv file that a user may provide to indicate additional metadata (ie. sample species, condition, description, replicate, etc.). This file can be associated with a plate during plate addition or from the Plates page to an existing plate.
From the Analyses page, select a completed analysis and then select the ‘Group Analysis’ tab to review the quantitation results organized in groups according to the sample metadata. From this page, select the ‘Setup Group Analysis’ button to access a guided wizard that will setup the differential expression analysis of specific samples or groups of samples.
Proteograph assay controls and their analysis results are tracked separately on the ‘QC Charts’ tab in addition to the other results tabs. From this page key performance metrics are automatically recorded and can be quickly reviewed relative to control results from other plates or studies. Statistical limits are calculated and displayed to evaluate deviations from expected ranges.
Analysis results can be exported by first selecting an analysis from the Analyses page, and then selecting the ‘Protein Group/Peptide Results’ button in the results tab. Results are exported as peptide and protein group .csv files.
The NP tables summarize LFQ results at the individual nanoparticle level. In the Panel tables, the single quantification value for a particular peptide or protein group represents the quantitation value of the NP measured most frequently across all samples (Max Representation roll-up)