The mAbXmise range of kits are designed to facilitate the implementation in the clinical lab of Promise technology for quantifying monoclonal antibodies (mAbs).
mAbXmise kits are ready-to-use kits, containing everything required to prepare the patient sample until the LC-MS injection. Interestingly, calibrators solutions and QCs (quality control) solutions are provided, thus avoiding the user to spend time in their preparation before each experiment. Also, the SIL-mAbs provided in mAbXmise kit are coated onto the sample preparation plate (see kit content). The patient samples will be then deposited on each well. The addition of the plasma sample will allow the resuspension of the SIL-mAbs. Doing so, the risk of error is limited. The same quantity of SIL-mAbs is loaded on each well, including wells for calibrators and QC solutions.
The range developed by Promise Proteomics includes two products:
- A kit specifically designed for monitoring therapeutic antibodies prescribed for inflammatory diseases
- A second kit, targeting antibodies prescribed for the treatment of cancer.
Tutorial Video mAbXmise Kits:
Quick Start: Starter KIT
Starter KIT use
The Starter KIT is a ready-to-use lyophilized solution containing the 9 mAbs of the 2 pre-digested KITs, a Belatacept Starter KIT is also available.. This solution has several objectives:
- Implement the LC-MS/MS method.
- Optimise the method parameters (source parameters, MS/MS, and chromatographic separation).
Control sample for the beginning and end of the analysis.
Preparation of the SK: Take up the freeze-dried residue with 400 µL H2O/ ACN (95/5%) (V/V). Allow to stand for 5 min then vortex vigorously for 30 seconds. Transfer the contents into 2 vials with insert (glass, avoid plastic containers as much as possible for adsorption of compounds).
The solution is stable for several weeks at room temperature for future analysis.
mAbXmise KIT: ITDM & OTDM multiplexed analysis
The gradient shown here is a first gradient for use with the StarterKIT. It should normally provide sufficient separation for proper analysis. In addition, it is intended to be adjusted according to the needs of the laboratory. If the analysis is suitable for the intended use, you can then fix it as the method gradient.
MS Target Scan Time
The final method should be Scheduled (Retention Time Window). The first analyses are advised to be without windows in order to fix the retention times, but as soon as the final gradient is fixed, retentions time windows are essential.
Generally, the width of the chromatographic peaks will be between 4 and 8 s (depending on the LC and columns). The objective is therefore to have a cycle time of 0.3 to 0.4 s and MRM windows of 24/30 s (depending on the reliability of the LC chain).
It is preferable to first carry out several analyses with the Starter KIT to check that the method works. If possible, send us the results (raw file) so that we can check that the method is workable.
For greater robustness and sensitivity of the analysis, it is necessary to use the whole set of transitions (Sum) in the method processing to quantify the analytes. Normally, for each mAb, 4 transitions will be monitored.
|Method Sciex – 5500 / 6500+ [OTDM-ITDM]||Download file here|
|Method ThermoFisher – Altis [OTDM-ITDM]||Download file here|
|Method ThermoFisher – Quantis [OTDM-ITDM]||Download file here|
|Method ThermoFisher – Exploris HR [OTDM-ITDM]||Download file here|
|Method Waters – Xevo TQD/TQS [OTDM-ITDM]||Download file here|
|Method Waters – Xevo TQXS [OTDM-ITDM]||Download file here|
|Method Shimadzu – TQ 8050/8060 [OTDM-ITDM]||Download file here|
- The column must be a C18 column, regardless of the supplier.
- The length of the column should be between 50 (Best Kind) and 100 mm.
- Internal diameter of 2.1 mm. An internal diameter of 1 mm should be avoided.
- Particle size between 1.5 and 2.5µm.
- Column temperature: depending on care and use, 40-50 °C.
For more information on Promise Proteomics, click here.