| Metadata |
| datasetIdentifier | PASS00853 |
| datasetType | SRM |
| submitter | Sofia Waldemarson <sofia.waldemarson@immun.lth.se> |
| submitter_organization | Immunotechnology Lund University |
| lab_head_full_name | Peter James |
| lab_head_email | peter.james@immun.lth.se |
| lab_head_organization | Immunotechnology Lund University |
| lab_head_country | Sweden |
| datasetTag | AFFIRM_3bead_to_pg |
| datasetTitle | Targeted measurement of proteins in serum in pg/ml range in the immunoaffinity platform AFFIRM |
| publicReleaseDate | 2016-12-01 00:00:00 |
| finalizedDate | 2016-03-30 01:43:26 |
| summary | This dataset demonstrates how oriented antibody immobilization to magnetic beads in the immunoaffinity platform AFFIRM greatly improves the detection capacity of the assay down to pg/ml range and with an improved multiplexing capacity with an 11-plex setup.
|
| contributors | Anna Säll, Daniel Corbee, Sara Vikström, Filip Ottosson, Helena Persson, Sofia Waldemarson |
| publication | Säll A, Corbee D, Vikström S, Ottosson F, Persson H, Waldemarson S - submitted manuscript |
| growth | not applicable |
| treatment | Samples are recombinant proteins spiked into a serum background at concentrations ranging between 100 ng/ml down to 50 pg/ml concentration. |
| extraction | Proteins are captured using recombinant scFv antibody fragments immobilized to magnetic beads. |
| separation | Upfront the SRM analysis the peptides are separated with an online HPLC-system using a 30 min gradient. |
| digestion | After washing, the proteins are digested directly sitting on the scFv on the beads 37 degrees over night. The resulting peptides are analyzed using SRM. |
| acquisition | The TSQ instrument was operated in positive ion mode with a spray voltage of 1.8 kV and an ion capillary temperature of 270 °C. Q1 and Q3 were set to unit resolution (0.7 Da) and analysis run with 10 ms dwell time. Data acquisition was done using Xcalibur software (version 2.1). |
| informatics | Raw files were converted into mzML format using the Proteios tools. |
| instruments | TSQ Vantage |
| species | Human |
| massModifications | none |
