Feedback

Robust Preanalytical Performance of Soluble PD-1, PD-L1 and PD-L2 Assessed by Sensitive ELISAs in Blood

Affiliation
German Heart Center Munich, Clinics at the Technical University Munich, Institute of Laboratory Medicine, Munich Biomarker Research Center, 80636 Munich, Germany
Krueger, Kimberly;
Affiliation
German Heart Center Munich, Clinics at the Technical University Munich, Institute of Laboratory Medicine, Munich Biomarker Research Center, 80636 Munich, Germany
Mayer, Zsuzsanna;
Affiliation
German Heart Center Munich, Clinics at the Technical University Munich, Department of Cardiovascular Disease, 80636 Munich, Germany
Kottmaier, Marc;
Affiliation
Department of Internal Medicine III, University Hospital Munich-Grosshadern, 81377 Munich, Germany
Gerckens, Miriam;
ORCID
0000-0002-1922-2127
Affiliation
Department of Internal Medicine III, University Hospital Munich-Grosshadern, 81377 Munich, Germany
Boeck, Stefan;
Affiliation
Department of Clinical Chemistry, University Hospital of the Technical University Munich, 81675 Munich, Germany
Luppa, Peter;
Affiliation
German Heart Center Munich, Clinics at the Technical University Munich, Institute of Laboratory Medicine, Munich Biomarker Research Center, 80636 Munich, Germany
Holdenrieder, Stefan

The interaction between programmed death-1 receptor PD-1 and its ligands PD-L1 and PD-L2 is involved in self-tolerance, immune escape of cancer, cardiovascular diseases, and COVID-19. As blood-based protein markers they bear great potential to improve oncoimmunology research and monitoring of anti-cancer immunotherapy. A variety of preanalytical conditions were tested to assure high quality plasma sample measurements: (i) different time intervals and storage temperatures before and after blood centrifugation; (ii) fresh samples and repeated freeze–thaw-cycles; (iii) different conditions of sample preparation before measurement. Concerning short-term stability, acceptable recoveries for PD-1 between 80 and 120% were obtained when samples were kept up to 24 h at 4 and 25 °C before and after blood centrifugation. Similarly, recoveries for PD-L2 were acceptable for 24 h at 4 °C and 6 h at 25 °C before blood centrifugation and up to 24 h at 4 and 25 °C after centrifugation. Variations for PD-L1 were somewhat higher, however, at very low signal levels. Sample concentrations (ng/mL) were neither affected by the freezing process nor by repeated freeze–thaw cycles with coefficients of variation for PD-1: 9.1%, PD-L1 6.8%, and PD-L2 4.8%. All three biomarkers showed good stability regarding preanalytic conditions of sample handling enabling reliable and reproducible quantification in oncoimmunology research and clinical settings of anti-cancer immunotherapy.

Cite

Citation style:
Could not load citation form.

Rights

License Holder: © 2022 by the authors.

Use and reproduction: