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Quantification of cleaved beta2-microglobulin in serum from patients undergoing chronic hemodialysis
Dorthe B Corlin1, Jette W Sen, Søren Ladefoged
1Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark.
Background:
Patients on chronic hemodialysis are prone to develop amyloid deposits of misfolded beta(2)-microglobulin (beta(2)M) in osteoarticular tissues. beta(2)M with various deletions/truncations and chemical modifications has been found together with structurally intact beta(2)M in extracts of beta(2)M amyloid fibrils. The state of the circulating population of beta(2)M molecules has not been characterized previously with high-resolution methods.
Methods:
We used immunoaffinity-liquid chromatography-mass spectrometry analysis of serum samples to examine whether structurally modified beta(2)M is generated in the circulation. In addition, we developed an immunoassay for the quantification of a cleaved beta(2)M variant in biological fluids based on novel monoclonal antibodies and applied this assay to patient and control sera.
Results:
A specific alteration compatible with the generation of lysine-58-cleaved and truncated beta(2)M (DeltaK58-beta(2)M) was found in the sera of many (20%-40%) dialysis patients but not in control sera or sera from patients with cerebral amyloidosis (Alzheimer disease). Applied to patient sera, specific immunoassays revealed that dialysis, as expected, significantly lowered the total beta(2)M concentration, but the concentrations of DeltaK58-beta(2)M remained unchanged after dialysis. The results also show that patients dialyzed with less biocompatible membranes have higher serum concentrations of cleaved beta(2)M (mean, 8.5, 1.8, and 0.7 mg/L in cuprophane membrane-dialyzed, polysulfone membrane-dialyzed, and control sera, respectively).
Conclusions:
This study for the first time demonstrates and assigns the structure of a specific beta(2)M variant in sera from dialysis patients. Because this variant is conformationally unstable in vitro, it may be involved in in vivo amyloidogenesis.
Insights
Dialysis patients develop a specific cleaved beta(2)-microglobulin (beta(2)M) variant in their circulation. This variant, linked to amyloid deposits, is not cleared by dialysis and is higher with less biocompatible membranes.
Area of Science:
- Biochemistry
- Nephrology
- Proteomics
Background:
- Patients undergoing chronic hemodialysis are susceptible to beta(2)-microglobulin (beta(2)M) amyloid deposits in osteoarticular tissues.
- Beta(2)M amyloid fibrils contain structurally intact and modified forms of beta(2)M, but the circulating beta(2)M population's state remains uncharacterized.
- High-resolution methods are needed to analyze the circulating beta(2)M population.
Purpose of the Study:
- To investigate the presence and structure of modified beta(2)M in the circulation of hemodialysis patients.
- To develop and apply a novel immunoassay for quantifying a specific cleaved beta(2)M variant.
Main Methods:
- Immunoaffinity-liquid chromatography-mass spectrometry (LC-MS) was used to analyze serum samples for modified beta(2)M.
- A novel immunoassay employing monoclonal antibodies was developed to quantify a cleaved beta(2)M variant in biological fluids.
- The immunoassay was applied to serum samples from dialysis patients and controls.
Main Results:
- A specific alteration, lysine-58-cleaved and truncated beta(2)M (DeltaK58-beta(2)M), was identified in 20%-40% of dialysis patient sera, but not in controls or Alzheimer's disease patients.
- Dialysis significantly reduced total beta(2)M but did not alter DeltaK58-beta(2)M concentrations.
- Patients dialyzed with less biocompatible membranes (cuprophane) showed significantly higher serum concentrations of cleaved beta(2)M compared to those using polysulfone membranes or controls.
Conclusions:
- This study provides the first structural characterization of a specific beta(2)M variant in the sera of dialysis patients.
- The identified DeltaK58-beta(2)M variant is conformationally unstable in vitro.
- The DeltaK58-beta(2)M variant may play a role in in vivo amyloidogenesis in hemodialysis patients.
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