Capillary Serum Protein Electrophoresis Research Articles

Effects of lipemia on capillary serum protein electrophoresis in native ultra-lipemic material and intravenous lipid emulsion added sera.

This study aims to investigate the effect of natural ultralipemic material (NULM) and intravenous lipid emulsion (IVLE) on capillary serum protein electrophoresis (SPEP). NULM material was prepared from leftover patients' lipemic serum sample (triglyceride concentration >2,000mg/dL) pool by a refrigerated high-speed centrifuge, and IVLE Omegaven lipid emulsion (30%) was used. Serum pools for interference study were prepared from patient samples for which serum protein electrophoresis was studied as Normal SPEP and M Peak SPEP. For both types of lipemia (DULM and IVLE), five pools with triglyceride concentrations of ∼4.52 mmol/L, ∼7.91 mmol/L, ∼14.69 mmol/L, ∼21.47 mmol/L, and ∼28.25mmol/L were prepared. SPEP was studied in each pool with Sebia Capillarys Minicap. A repeated measure ANOVA test was used to determine the difference between the pools, and interferograms were used to evaluate the interference effect. Interference was not detected in IVLE added Normal SPEP and M Peak SPEP pools, either % or concentrations of fractions. In NULM-added Normal SPEP and M Peak SPEP pools, significant positive interference in albumin % (p=0.002 and p<0.001 respectively) and significant negative interference in gamma% (p<0.001 and p=0.005 respectively) and M protein peak (p=0.002) fractions were detected. However, significant positive interference was seen only for albumin concentration fractions (p<0.001 for both pools). It is vital to use NULM instead of IVLE solutions in lipemia interference studies for all laboratory tests, including CZE SPEP. The fractions concentration values calculated with the total protein concentration should be used for evaluating SPEP results.

Laboratory characteristics of IgG4-related disease: A retrospective study from a single tertiary medical center.

Immunoglobulin G4-related disease (IgG4-RD) is an immune-mediated fibroinflammatory condition with unique histopathological features that can affect most organs, making diagnosis challenging. This study characterized detailed laboratory characteristics of IgG4-RD. Baseline clinical and laboratory features of 33 patients with IgG4-RD were reviewed, including serum IgG4 concentrations, serum free light chains (sFLCs), IgGĸ- and IgGλ-heavy/light chains (HLCs), capillary serum protein electrophoresis (SPE), and immunofixation electrophoresis (IFE) of IgG4 subclass. The cohort of 33 patients showed male predominance (94%), with 8 (24%) exhibiting multiple organ involvement. Most patients (88%) had an elevated IgG4 concentration, and 67% had elevated erythrocyte sedimentation rate and IgE levels. Median IgG4 concentration at baseline was significantly higher in patients with >2 organs involved than those with ≤2. Furthermore, erythrocyte sedimentation rate was significantly correlated with serum IgG4 concentrations at baseline. SPE results demonstrated polyclonal gammopathy in most patients. Half of the patients had an increased κ/λ sFLC ratio, 42% had an increased IgGκ/IgGλ HLC ratio. Most patients exhibited hypergammaglobulinemia in the anodal end of the ɤ region on SPE. This study describes detailed laboratory features of IgG4-RD. Although none of these tests are considered diagnostically sufficient by itself, the provided laboratory characteristics can increase awareness of this disorder and help distinguish it from other IgG4-RD mimics.

Qualitative dysalbuminemia in capillary serum protein electrophoresis: a case series of 70 patients

Capillary serum protein electrophoresis (CPE) is a high-resolution technique. Thus, it is highly sensitive to the detection of qualitatives dysalbuminemia. By the present study, we aim to describe modifications of albumin fraction within the electrophoretogramm and to expose the underlying diseases. Eight thousand and forty-two CPE were studied, among which 70 showed qualitative dysalbuminemia (frequency ≈1/100). The average age of the patients is 33years with a sex ratio (M / F) of 2.33. Quantitatively, the mean ±SD of albuminemia is 18.5 ±6.8g/L by turbidimetric assay and 20.8 ±7.0g/L by electrophoretic integration. Qualitatively, the albumin region presented different shapes: a simple thickening of the base, a shoulder to the right or to the left of the peak, one or more distinct additional peaks. The electrophoretic quantification of these modifications ranges from 1.8 to 11.2g/L. The most frequent causal diseases are nephrotic syndrome and cholestatic jaundice.

Reflexing Suspicious β-Region Findings on Capillary Serum Protein Electrophoresis to Immunofixation or Immunosubtraction for Detecting Monoclonal Proteins.

Monoclonal immunoglobulins (M-proteins) that migrate in the β region on serum protein electrophoresis (SPEP) are often cloaked by this region's normal constituents. The present study interrogates the utility of using both quantitative and qualitative alterations in β-region bands for detection of β-migrating M-proteins. Consecutive SPEP cases analyzed by capillary electrophoresis were searched to identify the initial workup on 1,841 patients with increased total β regions, suspicious β-region findings resulting in reflex immunofixation (IFE), or immunosubtraction (ISUB). To augment quantitative information, separate β1 and β2 measurements were established and retrospectively used to evaluate their sensitivity for M-protein detection. We identified M-proteins in 205 (11.1%) cases, including immunoglobulin A (IgA) (54%), IgG (24%), IgM (13%), and free light chain (9%) isotypes. Of the 15 cases flagged by separate β1 and β2 measurements that were not identified by total β-region measurement, 1 progressed to myeloma. Of the 56 β-migrating M-proteins identified by qualitative features but without increase in any of the β-region measurements, 1 progressed to myeloma. A combination of separate measurements for β1 and β2 regions together with detection of β-region distortions increase sensitivity for identifying β-migrating M-proteins via reflex IFE or ISUB.

Lorazepam causing drug-induced liver injury : Rare entity

IntroductionLorazepam is a benzodiazepine derivative that is globally used for the therapy of anxiety and insomnia.ObjectivesThe objective of our work was to show that Lorazepam can be a cause of unexpected liver injury even though it is a rare entity.MethodsWe reported the case of a patient who had a Drug-Induced Liver Injury (DILI) under Lorazepam. We performed a literature review based on a PubMed search with the following keywords: “Lorazepam,DILI”.ResultsA 20 year-old-Tunisian woman was hospitalized in the psychiatry department of the hospital of Nabeul in Tunisia for a brief psychotic episode.She had a DILI under Olanzapine, Chlorpromazine and Lorazepam, which conducted us to interrupt her treatments except for the Lorazepam(5mg/day). The hepatic tests went back to normal even under Lorazepam. Few days later, the liver enzymes increased again to reach very high levels. Extensive workup was negative for other causes of liver injury, including viral hepatitis A, B, C and E.; capillary electrophoresis of serum proteins was normal; Exhaustive immunological tests were performed searching for auto immune hepatitis(anti-smooth muscle antibodies, anti-LKM1, anti-LC1, anti-SLA/LP) primary biliary cholangitis(anti-mitochondrial antibodies, anti-GP210, anti-sp100) and other antibodies like antinuclear antibodies were negative. Liver biopsy showed polymorphic inflammatory infiltrate including some eosinophilic polynuclear cells and rare vaguely epitheloid macrophages, with necrotico-inflammatory foci in the lobules, all of which were consistent with DILI. Lorazepam was discontinued and within 10 days her liver enzymes decreased and completely normalized.ConclusionsLorazepam, with an unknown action mechanism, can be a cause of DILI.DisclosureNo significant relationships.

Diagnosing a Challenging Case of Primary Amyloidosis using Capillary Electrophoresis

Amyloid Light chain (AL) amyloidosis is characterised by deposition of intact free light chains or their fragments in extracellular space. Here, authors describe the journey of a diagnostically challenging patient who presented with features of nephrotic syndrome and was finally diagnosed with AL amyloidosis. A 45-year-old female presented to the Outpatient Department (OPD) with gradually progressive generalised body swelling. On examination, hepatomegaly, cardiomegaly and macroglossia were observed. Renal biopsy, capillary serum protein electrophoresis, immunotyping and serum free light chain assay were performed along with routine blood and urine investigations to detect the presence of monoclonal protein. Bone marrow biopsy was conducted for confirmation of diagnosis. Proteinuria with hypoalbuminaemia 1.8 g/dL was detected during routine investigations. Renal biopsy showed presence of amyloid deposits in glomerular mesangium and walls of medium sized blood vessels which tested positive for Immunoglobulin (Ig) G, IgA, kappa and lambda chains on immune florescence. Serum protein capillary electrophoresis findings demonstrated increase in beta 2 fraction and distortion in gamma region. Immuno typing showed presence of monoclonal IgA heavy chains and lambda light chains. Bone marrow biopsy confirmed presence of plasma cell dyscrasia. Based on these findings authors concluded that capillary gel electrophoresis is more sensitive method than agarose gel electrophoresis in detecting beta migrating monoclonal proteins.

Evaluation of the capillary electrophoresis of serum proteins on the automated Helena V8® system

Evaluation of the capillary electrophoresis of serum proteins on the automated Helena V8® system

Incidence Analysis of Monoclonal Gammopathy of Undetermined Significance in People over 40 Years of Age

To study the distribution of monoclonal gammopathy of undetermined significance(MGUS) in different age, sex and ethnic people over 40 years old. Five hundred and ninety-six people(over 40 years old) examened in the Health Examination center of the First Affiliated Hospital of Xinjiang Medical University from July 2017 to September 2017 were selected. Among 596 people, male 310, female 286, Han people 488, and Uygur ethnic people 108. According to age, 596 people were divided into 3 groups, (40-59 years old group, 60-79 years old group, over 80 years old group). First, all samples were screened by capillary serum protein electrophoresis. If the suspected monoclonal bands were found in the electrophoretogram, and then the specific protein types were determined by serum immunofixation electrophoresis. The total incidence of MGUS in 596 screened population was 4.027%. The incidence of MGUS in 40-59 years old group, 60-69 years old group and over 80 years old group were 1.762%, 2.929% and 10% respectively, and the differences among the groups were statistically significant(P<0.05). The incidence of MGUS in male (5.806%) was significantly higher than that in female (2.097%)(χ2=5.177，P<0.05). Binary Logistic regression analysis showed that over 80 years old and male were independent risk factors for MGUS(P=0.001, OR=4.188, 95%CI: 1.814-9.673, P=0.048, OR=2.605, 95%CI: 1.009-6.725). The types of immunoglobulin in patients with MGUS were mostly IgG, IgG(66.7%) was significantly more than IgA (29.2%)(χ2=21.375，P<0.05)，and there was no significant difference in the incidence of MGUS between people with in Kappa and Lambda. The age increase and male may increase the incidence of MGUS, the IgG is the most common type of immunoglobcdin in pathogenesis of MGUS, so the early screening should be done.

Utility of Reflexing Suspicious Beta Region Findings on Capillary Serum Protein Electrophoresis to Immunofixation for Detecting Monoclonal Gammopathies

Serum protein electrophoresis (SPEP) detects alterations suspicious for monoclonal immunoglobulins (M-protein/MP). Immunofixation (IFE) confirms/characterizes MPs. Beta region (BR) proteins, transferrin, and C3 can obscure beta-migrating MPs. There is no standardization of how to handle BR abnormalities. This study evaluates the effectiveness of our laboratory’s strategy of reflexing BR abnormalities to IFE and of separate measurement of beta1 and beta2 proteins for MP detection. In total, 2,134 SPEPs, performed on Sebia Capillarys, containing the comment “possible restriction in the beta region” (PRB) were identified by retrospective review (Michigan Medicine, 6/2013–6/2017). PRB was used in three situations: suspicious BR spike, beta1-beta2 bridge, or increased total beta region (TB) without obvious explanation (beta-gamma bridge), resulting in reflexed IFE. Inclusion of only the initial sample for each patient and exclusion of patients with MP history resulted in 1,904 cases. TB, MP, isotype, and increased polyclonal immunoglobulins were recorded. Separate beta1 and beta2 measurements were performed. Construction of references ranges (RRs) (mean ± 2 SD) using 31 normal patients showed RR 0.31 to 0.61 g/dL (beta1) and 0.24 to 0.57 g/dL (beta2). Normal TB RR, 0.65 to 1.03 g/dL. A total of 272 cases had MP detected by IFE (14.3%); 928 cases had increased TB (48.7%), while 660 had increased beta1/beta2 (34.7%). MP was present in 169 cases with increased TB (8.88%) and 183 with increased beta1/beta2 (9.61%). Low/normal TB, beta1, beta2, and beta1/beta2 were present in 103 (5.41%), 234 (12.3%), 119 (6.25%), and 88 (4.62%) cases with MP, respectively. Most common isotypes were IgA kappa and IgA lambda. Most common polyclonal increase was IgA (316). This technique had a true-positive rate of 14.3% with many negative IFEs, similar to another study reporting a 10% detection rate for elevated beta-fraction. Separate beta1 and beta2 measurements modestly increased MP detection. Further improvement of true-positive rates will require additional exclusion criteria.

Distortion of Capillary Serum Protein Electrophoresis Pattern May Indicate Monoclonal Gammopathy of Undetermined Significance but Not Multiple Myeloma

Distortion of Capillary Serum Protein Electrophoresis Pattern May Indicate Monoclonal Gammopathy of Undetermined Significance but Not Multiple Myeloma

Serum protein electrophoretic pattern in one-humped camels (Camelus dromedarius) in Tripoli, Libya.

The aim of this study was to characterize serum protein capillary electrophoretic pattern in apparently healthy adult male (age: 3-7 years) dromedary camels and also evaluate total protein and albumin levels using automated analyzer. Blood samples were taken from 20 camels. 5ml of blood was collected from the jugular vein and serum was separated from samples by centrifugation. Capillary electrophoresis of serum proteins identified six protein fractions in adult camels, including albumin, alpha1, alpha2, beta1, beta2 and gamma globulins, serum levels of these parameters were 3.9±0.04 g/dl, 0.16±0.01 g/dl, 0.39±0.03 g/dl, 0.515±0.03 g/dl, 0.205±0.01 g/dl and 0.61±0.04 g/dl, and 65.42±0.62 g/l, respectively. The total protein concentration was 65.42±0.62 g/L, while, the albumin/globulin (A/G) ratio was 2.4±0.14. The present study indicates six peaks with minicapillary electrophoresis and the results obtained were compared and interpreted in the light of finding reported by other investigators in camels.

Detection of Polyclonal Increases in Immunoglobulin G4 Subclass by Distinct Patterns on Capillary Serum Protein Electrophoresis: Diagnostic Pitfalls and Clinical Observations in a Study of 303 Cases.

Autoimmunity, hypersensitivity, and the recently recognized set of syndromes collectively termed immunoglobulin G4-related disease (IgG4-RD) may be associated with increased serum IgG4 levels. We reviewed our experience detecting increased IgG4 by distinct serum protein electrophoresis (SPEP) patterns. We studied 303 capillary SPEP cases with dome-like anodal γ changes and increased measured serum IgG4. IgG4 ranged from 208 to 6,670 mg/dL (normal, <201 mg/dL). Seventeen of 91 cases evaluated by immunosubtraction appeared monotypic (16 κ, 1 λ), but all five cases further analyzed by isoelectric focusing appeared polyclonal. Six cases with markedly increased IgG4 had presumptive evidence of IgG4-RD. Sixteen of 45 assessed patients had autoantibodies. Increased polyclonal IgG4 has a characteristic SPEP pattern that may mimic monoclonal gammopathy, even on immunosubtraction. κ Pseudo-restriction might reflect the naturally high κ/λ ratio of the IgG4 subclass. Autoantibodies were common, and the greatest IgG4 increases had clinical features of IgG4-RD.

Interferencia en electroforesis capilar debida a contraste yodado

Capillary electrophoresis of serum proteins is the method most currently used by clinical laboratories for the detection of monoclonal components in serum due to it being highly automated and rapid, with a high sensitivity, significantly improving the results compared to methods based on the use of gel techniques. However, there have been reports of interference by exogenous, non-protein substances that absorb at the same wavelength and cause false positives.Blood samples for laboratory analysis, including protein electrophoresis, were taken from two patients just after they had been subjected to a computed tomography (CT) scan with iodinated contrast. A monoclonal peak was observed in the beta fraction of the electrophoresis pattern, which was unable to be confirmed by immunofixation or immunosubtraction, leading to the suspicion of interference by iodinated contrast. A second sample was requested in both cases to repeat the analysis, which showed the disappearance of the peak, with a totally normal electrophoretic profile being obtained.

Does bilirubin interfere with capillary electrophoresis of serum proteins?

Capillary electrophoresis of serum proteins is a fast, reliable and simple technique, but many interference exist. The objective of our work is to study the interference of bilirubin on this technique; 70 icteric sera were analysed on Capillarys ™ (Sebia). A second electrophoresis was performed on 40 samples after bilirubin photodegradation. The bilirubin and serum proteins were determinated respectively by Jendrassik and Grof and biuret methods on Konélab 20i ™ (Thermo Electron Corporation). We found abnormal spreading of the albumin fraction of the anode side wich constitute sometimes an isolated fraction in the traditional area of pre-albumin migration. This fraction varies from 2.0 ± 2.0% (0.0 to 7.3%) or 0.98 ± 1.53 g/L (0 to 5.3 g/L) and it seems to be related to the direct bilirubin since, following overloading sera with a solution of bilirubin, no further fraction was recovered. An average decrease of bilirubin after photodegradation of 58 ± 17% (26-89%) is followed by a decrease in the same order 64 ± 38% (10-100%) of the additional fraction. Acetate cellulose electrophoresis of the same samples showed no variation. The high bilirubin levels seem modify slightly the electrophoretic profile. However the impact of the interference on the interpretation of electrophoretic trace is negligible.

Overlapping Presence of Macroamylasemia and Hyperamylasemia in Acute Pancreatitis

Overlapping Presence of Macroamylasemia and Hyperamylasemia in Acute Pancreatitis

Abstract 5557: Quantitative mass spectrometry assays for antibodies to measure multiple myeloma tumor burden and detect relapse

Abstract a) The goal of this study is to develop and characterize mass spectrometry assays for antibody quantification to assess multiple myeloma patients; the results generated from these peptide-based techniques will be compared data generated with gel-based and capillary serum protein electrophoresis (SPEP), which represents the current standard of care. b) Monoclonal antibody concentrations are currently determined using protein electrophoresis with either serum aliquots (SPEP) or Urine samples (UPEP), the specific class of myeloma is then determined using immunofixation. Liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM) assays have been developed to quantify all antibodies (Ig G, A, M, D, E, kappa, and lambda) and their isoforms in serum samples by measuring against known concentrations of stable isotope labeled internal standards using a triple quadrupole mass spectrometer. For assay development, at least two peptides representing each antibody isoform were derived from either LC-MS/MS runs based on excised gamma bands from stored SPEP gels of previous myeloma patients; these experimental results could also be supplemented by in silico workup of the protein sequence (i.e. theoretical peptide predictions). For each peptide target, several transitions (pairs of intact peptide masses and fragment ion masses) are programmed into the triple quadrupole for targeted detection using LC-MRM. Samples are analyzed in triplicate and compared with SPEP results. c) LC-MRM analysis has been performed on more than 40 serum samples collected from multiple myeloma patients. Using the selected peptides, protein amounts were calculated and compared with previous clinical SPEP results. The panel of antibodies presents unique opportunities and challenges for examining the utility of LC-MRM monitoring; for example, IgD and IgE increases can be detected with LC-MRM well below the 1 mg/ml limit of SPEP (at approximately 20 micrograms/ml). On the other hand, quantification of IgG1 with LC-MRM must be extremely accurate (&amp;lt; 10% CV) to detect the monoclonal protein against the background of that protein normally found in serum. To date, LC-MRM can detect all SPEP positive patients, and this method shows promise in detecting increases in antibody concentrations before they can be detected with SPEP. d) Quantitative mass spectrometry measurements for the antibodies combine the value of current SPEP and immunofixation experiments. Our results show that peptide-based monitoring can be as effective as protein-based monitoring. In addition, LC-MRM can detect light chain only disease, IgA myeloma that do not present a clear band in the gamma region of SPEP, and oligoclonal or polyclonal disease. These data enable broader commentary on the utility of LC-MRM in clinical protein measurements. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5557.

Unusual Serum Electrophoresis Pattern in a Woman with Pancreatic Carcinoma

A 51-year-old woman presented with progressive severe icterus associated with advanced inoperable pancreatic carcinoma. Biopsy of the liver revealed metastasis of a moderately differentiated adenocarcinoma, possibly of pancreatic origin. Imaging studies did not show any primary tumor in the pancreas but did reveal a deep vein thrombosis of the left lower extremity and a pulmonary embolism. Bone scans revealed metastases. The patient began treatment with chemotherapy (gemcitabine), radiotherapy to alleviate pain, and placement of a percutaneous transhepatic stent in the right biliary system because of progressive icterus. Routine biochemical investigation of the patient’s serum revealed increased total bilirubin (275 μmol/L, reference value <22 μmol/L), which consisted mainly of direct bilirubin (215 μmol/L, reference value <4 μmol/L). The cytosolic liver enzymes alanine aminotransferase (115 U/L) and aspartate aminotransferase (143 U/L) were moderately increased. The serum protein concentration was 68 g/L (reference interval 60–85 g/L), but capillary electrophoresis of serum proteins (CAPILLARYS 2, Sebia) demonstrated a low albumin fraction together with a marked additional peak observed between the β and the γ region (Fig. 1⇓ ). Results of immunofixation with antibodies against G, M, and A heavy chains and κ and λ light chains were negative, indicating that this additional electrophoretic fraction did not indicate the presence of a paraprotein. Agarose gel electrophoresis confirmed the abnormal pattern, ruling out analytical interference caused by atypical ultraviolet absorbance. Figure 1. Capillary electrophoresis of the patient’s serum. The arrow points to a marked additional fraction present between the β and γ zone. Absorbance is expressed as ultraviolet absorption …

Capillary zone electrophoresis of serum proteins: study of separation variables

Electrophoresis of serum proteins is one of the traditional applications of zone electrophoresis. Whereas electrophoresis in supporting media uses usually 5,5′-diethylbarbiturate at pH 8.6 as the buffer, in capillary zone electrophoresis with on-line UV detection, this electrolyte is of little use because of its high UV absorbance. For that reason, a number of operational electrolytes differing in composition were tested for use in capillary electrophoresis of serum proteins. The influence of the p K A of co-ions and counter ionsand the concentration of the operational electrolyte was examined. If 0.1 M methylglucamine-0.1 M ϵ-aminocaproic acid or 0.1 M methylglucamine, −0.1 γ-aminobutyric acid is used as the operational electrolyte, capillary electrophoresis separates serum proteins into more than ten zones.