doi: 10.1002/hem3.70053.
eCollection 2024 Dec.
Analysis of smoldering multiple myeloma according to the target of the monoclonal immunoglobulin of patients
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- PMID: 39670186
- PMCID: PMC11635023
- DOI: 10.1002/hem3.70053
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Analysis of smoldering multiple myeloma according to the target of the monoclonal immunoglobulin of patients
Hemasphere.
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No abstract available
Conflict of interest statement
Sylvie Hermouet, Nicolas Mennesson, Sophie Allain‐Maillet, Edith Bigot‐Corbel, Andri Olafsson, Brynjar Viðarsson, Páll T. Önundarson, Bjarni A. Agnarsson, Margrét Sigurðardóttir, Ingunn Þorsteinsdóttir, Ísleifur Ólafsson, Elías Eyþórsson, Ásbjörn Jónsson, Thorvardur J. Love, Saemundur Rognvaldsson, Einar S. Björnsson, Sigrún Thorsteinsdóttir, and Sigurdur Y. Kristinsson declare that they have no conflict of interest and nothing to disclose.International Myeloma Foundation.
Figures
Identified targets of purified monoclonal Ig from smoldering multiple myeloma (SMM) patients. (A) GlcSph immunoblot assays of SMM patients with a GlcSph‐reactive mcIg. For each patient, results were obtained in parallel with the unseparated serum IgG or IgA (in green) and the patient's purified mcIgG or mcIgA (in green). Measurement of Ig concentration, separation of the mcIg from other Igs by serum protein electrophoresis (SPE), and verification of the purity of the mcIg preparation by isoelectric focusing (IEF) were performed as described in the Supporting Information and published., , Polyvinylidene difluoride (PVDF) membranes were incubated in 100 μg/mL of GlcSph in 0.1 M sodium bicarbonate, rinsed three times, and then blocked for 2 h with 5% bovine serum albumin in phosphate‐buffered saline (PBS) and 0.1% Tween 20. Samples of purified mcIg were submitted to agarose gel electrophoresis, then the gels were blotted onto the GlcSph‐saturated membranes by diffusion blotting for 12 min., , After blocking for 1 h with 2.5% BSA in PBS and 0.1% Tween 20, membranes were incubated with peroxidase‐conjugated AffiniPure donkey antihuman IgG (H+L) antibody (Jackson ImmunoResearch) or horseradish peroxidase (HRP)‐conjugated goat anti‐human IgA α chain antibody (Bethyl Laboratories) for 1 h, then washed and revealed with Super Signal West Pico chemiluminescent substrate (Thermo Scientific). Signals corresponding to the patient's mcIg are encircled. The negative control is a patient with no reactivity for GlcSph. (B) Typical MIAA and dot blot assays of SMM patients with a monoclonal Ig that targets EBV. For each patient, results were obtained in parallel with the unseparated serum IgG or IgA (Serum, left) and the patient's purified mcIg (right) using the MIAA assay; results are shown as fluorescent intensity (FI). The FI values shown for each pathogen, Ag, protein, or lysate, were obtained after subtraction of the fluorescent background (B) of each pathogen protein or lysate. The thresholds of specific positivity, shown in dotted lines, were defined for each pathogen or protein: 1400 for EBV, CMV, VZV, HBV and B.burgdorferi, blue threshold; 1000 for HSV‐1 and HSV‐2, orange dotted line; 500 for HCV, H. pylori and T. gondii, green threshold. Dots may be superimposed; horizontal bars represent the means of results obtained for a pathogen (Ag, lysate). Experiments were performed in triplicate, and repeated at least once. Inserts show the results of EBV dot blot assays with recombinant EBV EBNA‐1 or VCA proteins; water was used as a negative control. Dot blot assays confirmed that the purified mcIg recognized EBV EBNA‐1 or VCA, as identified with the MIAA assay. Dot blot experiments were performed at least twice. (C) Typical MIAA assays of SMM patients with a monoclonal Ig that targets CMV. For each patient, results were obtained in parallel with the unseparated serum IgG or IgA (serum, left) and the patient's purified mcIg (right) using the MIAA assay; results are shown as FI. The FI values shown for each pathogen, Ag, protein, or lysate, were obtained after subtraction of the fluorescent background (B) of each pathogen protein or lysate. The thresholds of specific positivity (dotted lines) were defined for each pathogen or protein (1400 for CMV, blue threshold). Dots may be superimposed; horizontal bars represent the means of results obtained for a pathogen (Ag, lysate). Experiments were performed in triplicate and repeated at least once. Parts B and C show typical results of SMM patients; the results of all patients are shown in Supporting Information S1: Figure 3.
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