Task 1. Selection of biomarkers and panel design
Objective: to select biomarkers/nucleic acid modifications with the highest diagnostic and prognostic value in CLL.
Analysis of existing results and preliminary data, as well as supplementary analyses of archival material.
Evaluation of the usefulness of selected DNA/RNA modifications as differentiating markers (CLL vs. individuals without CLL; CLL vs. MBL/SLL) and as predictors of progression risk.
Definition of preliminary panel specifications (range of biomarkers, biological material, quality requirements).
Task 2. Adaptation and optimization of UPLC-MS/MS methods
Objective: to transfer and adapt the developed quantitative UPLC-MS/MS methods to routine diagnostic laboratory conditions, with an emphasis on efficiency, repeatability, and the possibility of standardization in accordance with regulatory requirements.
Optimization of sample preparation and chromatographic separation parameters and MRM detection.
Development of calibration and quality control (QC) procedures and validation parameters (precision, accuracy, LOQ/LOD, stability).
Workflow design with a view to automating sample preparation and minimizing operator errors.
Task 3. Flow cytometry – development and standardization of procedures
Objective: to develop repeatable flow cytometry procedures, including intracellular staining for nucleic acid modifications, and to integrate fluorescence analysis with image evaluation (population verification, artifact elimination).
Selection of antibody panels and measurement parameters for the assessment of B lymphocyte phenotype and clonality.
Standardization of staining procedures (including stages requiring high-temperature incubation) with parallel quality control based on cell imaging.
Definition of criteria for rejecting aggregates and artifacts and preparation of data analysis rules (gating and morphometric metrics).
Task 4. Automation of sample preparation and workflow standardization
Objective: maximum automation of sample preparation stages (isolation, staining, dilution, transfer) and ensuring a complete process audit trail.
Implementation of the Andrew+ robot as an open automation platform (working with standard consumables from different manufacturers).
Mapping of manual procedures in the form of robotic protocols with parameter and repeatability control.
Management of protocols and execution documentation in the OneLab environment (versioning, logs, traceability).
Task 5. Validation, testing, and preparation for implementation + dissemination
Objective: evaluation of the developed solutions in laboratory conditions and preparation of results for dissemination an