scholarly journals Comparison of IgA Heavy/Light Chain Immunoassays with Capillary Zone Electrophoresis and Total Immunoglobulin Measurements

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5692-5692
Author(s):  
Lidia Gartcheva ◽  
Violeta Petkova ◽  
Keranka Dimitrova ◽  
Veronika Petkova ◽  
Penka Ganeva ◽  
...  
Keyword(s):  
Capillary Zone Electrophoresis ◽  
Light Chain ◽  
Research Funding ◽  
Reference Range ◽  
Residual Disease ◽  
Normal Reference Range ◽  
Normal Reference ◽  
Normal Ranges ◽  
Zone Electrophoresis ◽  
Capillary Zone

Abstract Introduction: Capillary zone electrophoresis (CZE) and measurement of total immunoglobulins (total Ig) are standard techniques for the identification and quantification of monoclonal immunoglobulins (M-Ig). Heavy/light chain (HLC) pair analysis allows discrimination between Igκ and Igλ and as a result allows measurement of both the monoclonal involved and polyclonal uninvolved HLC pair. We compared measurement of M-Ig by CZE and total Ig to HLC levels. Methods: 93 samples from 8 patients with IgA intact immunoglobulin multiple myeloma (MM) were analysed. M-Ig was measured using CZE Sebia Capillarys 2 system, total IgA (tIgA) and IgAκ and IgAλ HLC concentrations on a SPAPLUS analyser. IgA HLC levels were measured with Hevylite® and compared to published normal ranges (IgAκ (g/L): 0.57-2.08, IgAλ (g/L): 0.44-2.04, IgAκ/IgAλ: 0.78-1.94). Passing and Bablok fit analysis was used to determine correlation between the assays. Results: Measurement of the involved HLC pair (iHLC) (median: 12.45g/L; range: 0.64-44.71g/L) compared well with CZE (n=34; median: 11.04g/L; range: 1.24-37.71g/L.; y= 1.2x + -2.65, R2= 0.94). Measurement of iHLC (median: 0.88. range: 0.05-21.55g/L) also compared well with tIgA measurement (n=65 median=1.44g/L; range: 0.227-21.11g/L, y=0.85x + -0.26, R2=0.98). Percent changes in iHLC concentrations from baseline through treatment compared well with CZE (n=28; y=1.59x + 0.15 R2=0.93) and tIgA (n=57; y=1.06x + 0.01, R2=0.96). Of the 34 samples with quantifiable M-protein by CZE, 32(94%) had an abnormal HLC ratio. The two discrepant samples were follow up samples from the same patient, where HLC normalised alongside total IgA entering the normal reference range. In addition, all 15 samples (15/65; 23%) where tIgA concentration was above the normal reference range all had abnormal HLC ratios. M-Ig was not detected by CZE in 48/82 (57%) samples, 46/48 of the samples (96%) had a normalised HLC ratio. In 38/65 (58%) samples, tIgA concentrations were within the normal reference range, and 34 (90%) had a normalised HLC ratio. HLC ratio for all patient samples normalised in subsequent samples following treatment. Conclusion: The measurement of M-Ig is comparable between Hevylite® and both CZE and tIgA. The presented data indicate that Hevylite® is a more sensitive test for detecting residual disease and warrants prospective studies on larger cohorts of patients. Acknowledgments:This work was partially supported by the National Science Fund (D02-35/2009). Disclosures Guenova: Novartis Pharma Sevices Bulgaria: Consultancy, Research Funding, Speakers Bureau; Roche Bulgaria: Consultancy, Research Funding, Speakers Bureau; Amgen Bulgaria: Consultancy, Research Funding, Speakers Bureau; Sanofi-Aventis Bulgaria: Consultancy, Research Funding, Speakers Bureau.

scholarly journals A Novel Risk-Model to Predict Time to First Treatment (TTT) in Chronic Lymphocytic Leukemia Based on Heavy+Light Chain Immunoparesis and Serum Free Light Chain Analysis: Results from the Israeli CLL Study Group

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1849-1849
Author(s):  
Tamar Tadmor ◽  
Andrei Braester ◽  
Najib Dally ◽  
Ariel Aviv ◽  
Yair Herishanu ◽  
...  
Keyword(s):  
Binding Site ◽  
Light Chain ◽  
Research Funding ◽  
Reference Range ◽  
Lymphocytic Leukemia ◽  
Light Chains ◽  
Normal Reference Range ◽  
Site Group ◽  
Normal Reference ◽  
Time To First Treatment

Abstract Introduction: Chronic Lymphocytic Leukemia (CLL) is frequently accompanied by immune dysregulation. Hypogammaglobulinemia is one of the most important immune defects encountered, and all three classes of immunoglobulins (IgG, A and M) can be involved. Recently, novel heavy+light chain (HLC) immunoassays have become available that quantify the light chain types of each immunoglobulin class (e.g. IgGk and IgGl). These assays are measured in pairs and provide information on the isotype produced by a tumour (the "involved" HLC), the non-clonal ("uninvolved") HLC, and the ratio (e.g. IgGk/IgGl) - which indicates monoclonality. HLC assays have been shown to improve monitoring of plasma cell dyscrasias, but their role in CLL is yet to be studied. Methods This is a multi-center study performed in collaboration with the Israeli CLL Study Group and involved 10 medical centers. The cohort included 122 patients with CLL and 26 healthy controls. Baseline was defined as the time the blood sample was taken. Serum samples were analyzed for levels of IgG subclasses (IgG1, IgG2, IgG3, IgG4), heavy+light chains (HLC) (IgGκ, IgGλ, IgAκ, IgAλ, IgMκ or IgMλ) and free light chains (sFLC). HLC-pair suppression was defined as an abnormal HLC ratio and uninvolved HLC levels below the normal reference range (i.e. in g/L: IgGκ<3.84; IgGλ<1.91; IgAκ<0.57; IgAλ<0.44; IgMκ<0.19; IgMλ<0.12). HLC immunoparesis was defined as HLC isotype levels below the normal reference range, regardless of HLC ratio (i.e. HLC immunoparesis of at least 1 isotype indicates at least one of IgGκ, IgGλ, IgAκ, IgAλ, IgMκ or IgMλ below the normal reference range). Severe HLC-pair suppression or severe HLC immunoparesis was defined as a concentration of the uninvolved HLC or any HLC isotype that is suppressed by >50% below the normal reference range. The association of variables with time to first treatment (TTT) was performed with Cox proportional hazard model. Results Current analysis was performed on 105 CLL patients who had complete data available; median age was 68 years, 64% were males and Binet stage A, B, C, were 80%, 18.1% and 1.9% respectively. Median time from diagnosis to baseline measurements was 27 months (range 0-328 months). An abnormal sFLC ratio was identified in 70% (73/105) patients and summated k and λ concentrations (SFLC) were ≥70 mg/L in 18% of cases. An abnormal HLC ratio was present in 32% (34/105) patients, and 21% had HLC pair suppression of any 1 HLC isotype. HLC immunoparesis of 1, ≥2 and ≥3 isotypes was observed in 74 (70%), 58 (55%) and 36 (34%) of patients, respectively, with severe HLC immunoparesis identified in 40 patients (38%). Patients with IgG2 suppression were more frequently hospitalized due to infection with an Odds Ratio of 3.826 (p=0.031). In multivariate analysis, SFLC ≥70 mg/L and severe HLC immunoparesis were independently associated with TTT (HR 15.3, p<0.001; HR 80, p<0.001 respectively). Using these 2 variables, a risk-stratification model was constructed that separated CLL patients into 3 risk groups (with 0, 1 or 2 risk factors) with significantly different TTT (p<0.001, Figure 1). Patients with both risk factors (SFLC ≥70 mg/L and severe HLC immunoparesis) had the shortest TTT. Conclusions The findings presented here demonstrate that there is considerable potential for the use of HLC and FLC immunoassays to provide prognostic information in CLL. Figure 1. Figure 1. Disclosures Tadmor: PFIEZER: Consultancy; JNJ: Consultancy; ABBVIE: Consultancy; NOVARTIS: Consultancy; ROCHE: Research Funding. Aviv:ABBVIE: Consultancy; ROCHE: Research Funding. Herishanu:ROCHE: Research Funding; JNJ: Consultancy; ABBVIE: Consultancy. Shvidel:ROCHE: Consultancy, Research Funding; ABBVIE: Consultancy, Research Funding; JNJ: Consultancy. Rahimi-Levene:ABBVIE: Consultancy. Ruchlemer:ABBVIE: Consultancy; JNJ: Consultancy. Fogl:The Binding Site Group Ltd: Employment. Polliack:ROCHE: Research Funding; ABBVIE: Consultancy. Magal:The Binding Site: Employment. Townsend:The Binding Site Group Ltd: Employment.

scholarly journals A Method to Measure Voxelotor Exposure in People with Sickle Cell Disease Using Capillary Zone Electrophoresis

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 2025-2025
Author(s):  
Susanna A Curtis ◽  
Elana M Friedman ◽  
Caterina Minniti ◽  
Annie Ngyuen Dang ◽  
Mira Pochron ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Capillary Zone Electrophoresis ◽  
Sickle Cell ◽  
Whole Blood ◽  
Blood Concentration ◽  
Research Funding ◽  
Cell Disease ◽  
Calculated Concentration ◽  
Zone Electrophoresis ◽  
Capillary Zone

Abstract Background: Voxelotor (Oxbryta®) is a small molecule that binds to the alpha chain of hemoglobin (Hb) and increases the affinity of Hb for oxygen which reduces sickle Hb polymerization. It was approved by the FDA in 2019 for the treatment of sickle cell disease (SCD). Currently, the only method available to estimate the concentration of voxelotor in the blood is to obtain exposure measurements which are only available in select research laboratories. A method to measure voxelotor at a standard laboratory would allow clinicians to assess compliance and may be useful in determining optimal dosing. Case studies have reported that voxelotor binding to Hb interferes with capillary zone electrophoresis (CZE). As previously reported, in CZE the characteristic peaks of hemoglobin A2 (HbA2) and hemoglobin F (HbF) split in the presence of voxelotor. Interestingly, it does not cause the hemoglobin S peak to split. We posited that we could use this split to estimate the presence of voxelotor and whole blood concentration. Biophysical measurements of voxelotor binding to Hb were also measured in these samples. Methods: Patients were enrolled prospectively in an IRB approved protocol. Voxelotor was initiated at 1500 mg daily on day 0 and samples were taken at day 0 (pre-dose), 14, 30, and 60. Samples had Hb variants quantified by CZE using the Capillarys 2 FlexPiercing Instrument (Sebia, Georgia). Hematology parameters were measured with the Sysmex XN-1000 automated analyzer (Sysmex, Illinois). To determine whole blood concentration of voxelotor, samples were sent to Worldwide Clinical Trials where a validated liquid chromatography-tandem mass spectrometry method was used. Voxelotor's interference with HbA2 on CZE is dependent on the HbF percentage, therefore samples from patients with SCD were combined into three pooled samples (5-10 samples per pool) of HbF percentages spanning 5-30%. Three hundred µL aliquots of each pool were spiked with voxelotor in DMSO in triplicate to different concentrations between 0 and 600 µMol/L and were incubated at room temperature for 1 hour before being tested with CZE. Samples were then analyzed for voxelotor interference resulting in split peaks of HbF and HbA2. HbA2 interference percent (%VarA2) was calculated as the reported value of the HbA2 split peak over the total HbA2 value (both split and parent peak) times 100. F% was used as directly reported by the instrument without consideration of voxelotor interference. Results were then analyzed in Excel (Data Analytics package) using a multiparameter regression to generate a line of best fit. To allow for logarithmic fit when examining the correlation of calculated concentration with increase in Hb due to voxelotor, samples with negative Hb rises were excluded and concentrations which resulted as negative values were changed to 0.01 µM. Results: Of 20 patients which have been enrolled to date, 9 patients have completed the study and their data was used for these analyses. Using the CZE method described above the concentration of voxelotor was quantifiable using the following equation. Equation 1: uM voxelotor = -99.13 + 7.10*%HbF +12.52*%VarA2 The calculated concentrations of voxelotor based on CZE results had a strong correlation with whole blood concentration (R 2 = 0.85, p &lt;0.001). (Figure 1) When calculated concentration was compared to change in Hb at days 14, 30, and 60 there was a significant positive logarithmic correlation between concentration and change in Hb (R 2=.56, p&lt;0.01). (Figure 2) Conclusions: Using equation 1, CZE can be used to detect the presence of voxelotor and estimate its whole blood concentration. This will allow clinicians to have a better understanding of how their patients are using voxelotor. Additionally, higher calculated whole blood concentrations correlated with higher increases in Hb. It was previously shown that patients who receive higher doses of voxelotor have on average larger increases in Hb. If it could be shown that increasing concentration in an individual on voxelotor is associated with an increased Hb for that individual, then our method could also be used to help clinicians select and adjust doses of voxelotor in a similar manner to how HbF is used in hydroxyurea dosing. Figure 1 Figure 1. Disclosures Curtis: GBT: Consultancy. Minniti: CSL Behring: Other: Endpoint adjudicator; Bluebird Bio: Other: Endpoint adjudicator; F. Hoffmann-La Roche: Consultancy; Chiesi: Consultancy; Novo Nordisk: Consultancy; Forma: Consultancy; Novartis: Consultancy; GBT: Consultancy. Ngyuen Dang: GBT: Current Employment. Pochron: GBT: Current Employment. Campbell: GBT: Research Funding; Sebia: Research Funding.

Capillary zone electrophoresis for haemoglobinopathy diagnosis

2012 ◽  
Vol 66 (1) ◽  
pp. 29-39 ◽  
Author(s):  
Nicole Borbely ◽  
Lorraine Phelan ◽  
Richard Szydlo ◽  
Barbara Bain
Keyword(s):  
Capillary Electrophoresis ◽  
Capillary Zone Electrophoresis ◽  
High Performance ◽  
Reference Range ◽  
Specific Reference ◽  
Specific Method ◽  
Alkaline Ph ◽  
Zone Electrophoresis ◽  
Microcolumn Chromatography ◽  
Capillary Zone

AimsCapillary zone electrophoresis (CE) at alkaline pH is increasingly used in haemoglobinopathy diagnosis. We report our evaluation of automated CE, using the Capillarys 2 Flex Piercing instrument, as a routine diagnostic method for the detection of variant haemoglobins and the diagnosis of β thalassaemia.MethodsA Capillarys 2 Flex Piercing instrument with Phoresis software was evaluated in our laboratory over a 6-week period, comparisons being made with high performance liquid chromatography (HPLC) and, for haemoglobin A2 quantification, with microcolumn chromatography.ResultsThe instrument was easy to use and was suitable for the quantification of haemoglobin A2. Quantification of A2 was precise and the percentage was stable with ageing of the blood specimen. Results differ among HPLC, CE and microcolumn chromatography and use of an instrument-specific, method-specific reference range is therefore recommended until such time as there is standardisation between methods and manufacturers. Common variant haemoglobins were provisionally identified without difficulty. There are some uncommon variant haemoglobins that are detected by HPLC but not by capillary electrophoresis, but the reverse also occurs.ConclusionsCapillary electrophoresis using a Capillarys 2 Flex Piercing instrument is suitable for haemoglobinopathy diagnosis.

scholarly journals Detection of Monoclonal Proteins in Sera by Capillary Zone Electrophoresis and Free Light Chain Measurements

2002 ◽  
Vol 48 (9) ◽  
pp. 1600-1601 ◽  
Author(s):  
Godelieve Mariën ◽  
Els Oris ◽  
Arthur R Bradwell ◽  
Norbert Blanckaert ◽  
Xavier Bossuyt
Keyword(s):  
Capillary Zone Electrophoresis ◽  
Light Chain ◽  
Free Light Chain ◽  
Zone Electrophoresis ◽  
Capillary Zone ◽  
Monoclonal Proteins

scholarly journals Lifestyle Change Alone Sufficient to Lower Cholesterol in Male Patient With Moderately Elevated Cholesterol: A Case Report

2018 ◽  
Vol 13 (2) ◽  
pp. 148-155
Author(s):  
Walter J. Janse Van Rensburg
Keyword(s):  
Cardiovascular Disease ◽  
Case Report ◽  
Reference Range ◽  
Lifestyle Change ◽  
Cholesterol Lowering ◽  
Normal Reference Range ◽  
Cholesterol Levels ◽  
Normal Reference ◽  
Normal Ranges ◽  
Elevated Cholesterol

Background. Cardiovascular disease is a major cause of deaths. Elevated cholesterol levels to above the normal reference range is a major risk factor for developing cardiovascular disease. Current guidelines recommend the use of cholesterol-lowering drugs to lower cholesterol levels to within the normal reference range. However, the American Heart Association further recommends a change in lifestyle in managing cholesterol levels. Thus, cholesterol-lowering drugs may not be needed if a lifestyle-change alone is sufficient in lowering cholesterol levels to within normal ranges. Unfortunately, limited examples exist in academic literature to illustrate the effectiveness of lifestyle change alone in lowering of cholesterol levels. Case report. We report a case of a 33-year-old man, with moderately elevated cholesterol levels and a family history of cardiovascular disease. Method. The man followed an altered healthy fat diet accompanied with moderate exercise for 6 weeks, without the addition of cholesterol-lowering agents. Results. At the 6-week follow-up, he was able to decrease his total cholesterol by 40.25% and low-density lipid cholesterol by 52.8%, to within normal ranges. The cholesterol levels remained within normal ranges after 6 months. Conclusion. This case illustrates that in some individuals, lifestyle change alone is sufficient to lower moderately elevated cholesterol levels.

Evaluation of normal reference range for thyroid uptake of technetium-99 m in a single centre UK population

2017 ◽  
Author(s):  
Mavin Macauley ◽  
Mohamed Shagwi ◽  
Kim Howe ◽  
Andrew Curry ◽  
Elizabeth Howell ◽  
...  
Keyword(s):  
Reference Range ◽  
Single Centre ◽  
Normal Reference Range ◽  
Normal Reference ◽  
Thyroid Uptake
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