Assessment of the influence of the patient's inflammatory state on the accuracy of a haptoglobin selected reaction monitoring assay.
BACKGROUND
The use of targeted LC-MS/MS methods for protein quantitation in clinical laboratories implies a careful evaluation of potential sources of analytical interference. In this study, we investigated whether inflammation, which is associated with both the release of proteolytic enzymes and increased expression of acute phase protease inhibitors, is affecting the accuracy of a haptoglobin selected reaction monitoring (SRM) assay.


RESULTS
A SRM assay was developed and used to quantify haptoglobin in 57 human serum samples. The SRM assay had CVs (n = 6) of 12.9% at 698 mg/L and 11.8% at 1690 mg/L. Results of the SRM assay were compared to those of a commercial immunonephelometric test. Passing-Bablok regression gave a proportional bias of 0.92 (95% CI: 0.82 to 1.04) and a constant bias of 75.40 (95% CI: -71.09 to 251.04), indicating that SRM and immunonephelometric assays provided comparable results. We then investigated whether the accuracy of the SRM assay was influenced by the patient's inflammatory state by assessing the relationship between the serum CRP concentration and the bias between the two methods. No correlation was found between the SRM/immunoassay bias and the CRP concentration (Pearson correlation coefficient r = 0.0898).


CONCLUSIONS
These data indicate that neither the release of proteolytic enzymes nor the increased level of protease inhibitors occurring during inflammation processes have a significant impact on the haptoglobin SRM assay accuracy. Such studies provide important information about potential sources of analytical interferences in protein SRM assays.
Department of Genetic and Laboratory Medicine, Geneva University Hospitals, Geneva, Switzerland.
Olivier
O
Lassout
Department of Genetic and Laboratory Medicine, Geneva University Hospitals, Geneva, Switzerland ; Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Geneva, Switzerland.
Denis
D
Hochstrasser
Department of Genetic and Laboratory Medicine, Geneva University Hospitals, Geneva, Switzerland ; Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Geneva, Switzerland.
Pierre
P
Lescuyer
Haptoglobin; Inflammation; Protease inhibitor; Quantitative proteomics; Selected reaction monitoring; Trypsin digestion