scholarly journals Abnormal Heavy/Light Chain Ratio after Treatment Is a Robust Predictor of Shorter Survival in Patients with IgA Myeloma

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3254-3254
Author(s):  
Yoshiaki Abe ◽  
Yasuhito Suehara ◽  
Hiroyuki Takamatsu ◽  
Yoshiaki Usui ◽  
Kentaro Narita ◽  
...  
Keyword(s):  
Overall Survival ◽  
Serum Protein ◽  
Light Chain ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
M Protein ◽  
Lower Sensitivity ◽  
Prognostic Impact ◽  
Monoclonal Protein ◽  
Significant Difference

Abstract Accurate quantification of monoclonal protein (M-protein) is essential for response assessment, management, and prediction of prognosis in patients with multiple myeloma (MM). Serum protein electrophoresis (SPEP) is commonly used to determine the degree of M-protein reduction in intact immunoglobulin (Ig) MM. However, SPEP has limitations when M-protein comigrates into the beta-fraction or M-protein fails to show a distinct sharp spike on densitometry. Ig heavy/light chain (HLC) assay enables separate quantification of the different light chain types of each Ig class. Although HLC assay quantifies different light chain subtypes of Ig classes, the sensitivity for detecting M-protein clonality and its impact on outcome may differ between IgA and IgG myeloma. To investigate the clinical and prognostic impact of HLC assay, we retrospectively analyzed the correlation of heavy/light chain ratio (HLCR) with clinical status and its impact on outcome. A total of 402 frozen serum samples from 120 patients with MM (41 for IgA and 79 for IgG) treated at Kameda Medical Center (Kamogawa-shi, Chiba, Japan) and Kanazawa University Hospital (Kanazawa-shi, Ishikawa, Japan) at the times of various IMWG responses were collected. Samples were analyzed using the HLC assay, and the results were compared with serum protein electrophoresis (SPEP), free light chain ratio (FLCR), immunofixation, total IgG, IgA, and overall survival (OS). Percentages of samples with normal HLCR at presentation, PR, VGPR, CR, and sCR were 0%, 0%, 27.6%, 100%, and 88.9%, respectively, for IgA MM and 0%, 12.5%, 64.3%, 100%, and 84.3%, respectively, for IgG MM. Normalization of HLCR at VGPR was more frequent in IgG MM compared to IgA MM (PR; 12.5% vs. 0%, respectively, P = 0.169, VGPR; 64.3% vs. 27.6%, respectively, P = 0.004), which suggests the lower sensitivity of detecting clonality in patients with IgG MM than those with IgA MM. Abnormal HLCR was seen more frequently in patients with poorer myeloma response, and it appeared to be more sensitive for detecting clonality in IgA myeloma compared to IgG myeloma after treatment. No significant difference in OS was observed between patients with or without uninvolved Ig suppression and OS if they obtained ≥ VGPR. Among the 85 patients that achieved ≥ VGPR, those that remained HLCR abnormal showed significantly shorter overall survival (OS) compared to those achieving normal HLCR (not reached vs. 55.5 months, P = 0.032). This correlation was seen in IgA myeloma patients (not reached vs. 30.1 months, P = 0.014), but not in IgG myeloma patients when analyzed separately. Univariate and multivariate analyses of factors that may affect survival identified abnormal HLCR at the best response as the only independent risk factor (hazard ratio, 4.7; 95% confidence interval, 1.4 - 15.26; P = 0.012) for shorter OS in this subset of patients. In conclusion, HLC assay is useful for accurate monitoring of monoclonal protein in patients with myeloma. The results suggested that obtaining normal HLCR indicated a more favorable prognosis in patients with IgA myeloma, but not IgG myeloma, that achieved VGPR or better response. Disclosures Takamatsu: Celgene: Honoraria; Janssen Pharmaceuticals: Honoraria.

Serum Free Kappa to Free Lambda Ratios as an Adjunct to Serum Protein Electrophoresis for the Detection of Monoclonal Proteins in the Serum.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4856-4856
Author(s):  
Arthur R. Bradwell ◽  
Jean Garbincius ◽  
Earle W. Holmes
Keyword(s):  
Serum Protein ◽  
Light Chain ◽  
Protein Electrophoresis ◽  
Free Light Chain ◽  
Serum Protein Electrophoresis ◽  
Serum Free ◽  
Monoclonal Protein ◽  
Serum Free Light Chain ◽  
Monoclonal Proteins

Abstract Serum free light chain measurements have been shown to be useful in the diagnosis and monitoring of patients with monoclonal gammopathies. The present study was undertaken to evaluate the effect of adding the measurement of serum free light chain kappa to lambda ratios to the serum protein electrophoresis evaluation that we typically use as an initial screen for the detection of monoclonal proteins. We retrospectively tested 347 consecutive samples from individuals who had no previous history of plasma cell dyscrasia and had not previously had a serum or urine electrophoresis or immunofixation electrophoresis test at our institution. The quantitative serum protein electrophoresis test that was ordered was performed using Hydragel Beta 1- Beta 2 gels and Hydrasis instrument (Sebia, Inc., Norcross, GA). The protein content of the electrophoresis zones were quantitated by scanning densitometry and the electrophoresis pattern of each sample was qualitatively examined for abnormal bands and suspicious findings by a single, experienced observer. Serum free light chain concentrations and the serum free light chain kappa to lambda ratios were determined using the Freelite Human Kappa and Lambda Kits (The Binding Site Ltd, Birmingham, UK) and the Immage analyzer (Beckman Coulter Inc., Brea, CA). The serum free light chain kappa to lambda ratios were outside the reference interval (0.25 to1.65) in 23 of the samples. Ten abnormal ratios were observed among a group of 57 samples that had either positive or suspicious qualitative evaluations for the presence of a restriction or that demonstrated hypo-gammaglobulinemia. Both abnormalities led to recommendations for follow-up testing, which confirmed the presence of a monoclonal protein in 21 of the samples. Six abnormal ratios were observed among a group of 159 specimens that had quantitative abnormalities in albumin or one or more of globulin fractions (hypo-gammaglobulinemia excepted) and normal qualitative evaluations. Seven abnormal ratios were observed among a group of 131 samples that had normal quantitative results and normal qualitative evaluations. Follow-up testing is not usually recommended for serum protein electrophoresis results like those in the latter two groups. We found that the addition of the serum free light chain kappa to lambda ratio to the serum protein electrophoresis test increased the number of abnormal screens that would have required further clinical and/or laboratory evaluation by 23%(i.e. from 57 to 70). Given the high specificity of the serum free light chain kappa to lambda ratio for monoclonal light chains, the additional 13 abnormal samples identified by this test are expected to have a high likelihood of harboring a monoclonal protein that would have otherwise eluded detection. Pending a definitive prospective study, we estimate that the addition of a serum free light chain kappa to lambda ratio to the serum protein electrophoresis screen would increase the rate of detection of serum monoclonal proteins by as much as 1.6-fold.

scholarly journals Comparison of Kappa & Lambda Freelite to Total Kappa & Lambda Immunoassays for the Detection of Monoclonal Gammopathies, Both As Standalone Tests and Alongside Serum Protein Electrophoresis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5705-5705 ◽  
Author(s):  
Vania T M Hungria ◽  
Petros Kampanis ◽  
Mark T Drayson ◽  
Tim Plant ◽  
Edvan de Queiroz Crusoe ◽  
...  
Keyword(s):  
B Cell ◽  
Serum Protein ◽  
Light Chain ◽  
Protein Electrophoresis ◽  
Immunoglobulin M ◽  
Serum Protein Electrophoresis ◽  
Monoclonal Protein ◽  
Monoclonal Gammopathies ◽  
Total Light ◽  
Age Range

Abstract Introduction Monoclonal gammopathies are a disparate group of diseases from benign to malignant which are characterised by the proliferation of a single B cell clone that produces a homogeneous monoclonal immunoglobulin (M-Ig). The method of detection and quantification of the M-Ig will depend upon whether it is an intact immunoglobulin or present as serum free light chain only. Historically serum (SPEP) and urine (UPEP) electrophoresis were considered the gold standard for identifying intact M-Ig and FLC respectively. In 2001 the introduction of the Freelite test changed the diagnostic and monitoring paradigm. The assay is now recommended as a tool to diagnose and monitor patients with B cell disorders. However, the assay is sometimes confused with monospecific immunoassays for measuring total kappa and total lambda. Here we compare kappa & lambda Freelite with total kappa & lambda immunoassays alongside SPEP as tools to identify patients with monoclonal gammopathies. Materials and Methods Sera from 102 blood donors (55 males and 47 females, age range 18-67 years) and 103 patients with light chain associated gammopathies (44 males and 59 females, age range 38 to 88 years, 60 kappa / 43 lambda)taken during the course of their treatment were available. The sera was analysed retrospectively with FreeliteTM (The Binding Site Ltd, Birmingham, UK) on a SPAPLUSand Total Kappa &Lambda nephelometricassays (Beckman Coulter, USA) on an Immage.Monoclonality was identified by results falling outside of manufacturers normal ratio ranges (Freelite 0.256-1.65, Total light chain 1.53-3.29). Serum protein electrophoresis was performed and unexpectedly positive or negative results were assessed using immunofixation on the Hydrasys electrophoretic system (Sebia, France). Results Monoclonal production was identified in 80/103 light chain associated gammopathiesby Freelite, negative IFE confirmed the absence of monoclonal protein in 22/23 patients with normal FLC kappa/lambda ratios and 1/23 patients had an IgG lambda intact immunoglobulin. SPEP was positive in 30/103 patients, with total kappa/lambda immunoassays detecting monoclonal protein in just 26/103 samples. Freelite was positive in 6/102, SPEP in 2/102 and total kappa/lambda in 8/102 normal blood donor sera. Interestingly, in 1 patient with an abnormal FLC ratio and total kappa/lambda result had a lambda light chain identified using IFE.Comparisons between the performances of Freelite, Freelite + SPEP, Total kappa/lambda and total kappa/lambda + SPEP are shown in table 1). Conclusion In keeping with Kyle et al (1999) our study confirms the limitations of total kappa / lambda assays as tools to identify M-Igs. This is the first study looking to apply the recommended algorithm of Freelite + SPEP to the total kappa/lambda assays. The addition of SPEP to total kappa/lambda assays improved the performance to detect abnormalities, but even combined they were neither as sensitive, specific or accurate as the Freelite assay. Given the limitations of the total light chain assays identified in our study, it is important that physicians are aware of which assay is being utilised; one easy method to discriminate would be to look at the normal range of the assay being reported. Table 1: Comparison of Freelite, Freelite + SPEP, Total kappa/lambda, Total kappa/lambda + SPEP Freelite Freelite + SPEP Total Total + SPEP Sensitivity 77.67 81.55 25.24 43.69 Specificity 94.12 92.16 92.16 91.18 PPV 93.02 91.30 76.47 83.33 NPV 80.67 83.19 54.97 61.59 Accuracy 85.85 86.83 58.54 67.32 Disclosures No relevant conflicts of interest to declare.

scholarly journals Prevalence of Monoclonal Gammopathy of Undetermined Significance in a Large Population with Annual Physical Examination in Beijing, China

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5519-5519
Author(s):  
Jinuo Wang ◽  
Jian-Hua Han ◽  
Yue-lun Zhang ◽  
Xin-xin Cao ◽  
Dao-Bin Zhou ◽  
...  
Keyword(s):  
Physical Examination ◽  
Light Chain ◽  
Chinese Population ◽  
Monoclonal Gammopathy ◽  
Conflicts Of Interest ◽  
Large Population ◽  
M Protein ◽  
Monoclonal Protein ◽  
Significant Difference ◽  
Undetermined Significance

Introduction Monoclonal gammopathy of undetermined significance (MGUS) is a clinically asymptomatic premalignant plasma cell disorder. Previous studies in Western countries have described the prevalence of MGUS in Caucasians. However, data is limited in Chinese population. We therefore performed this study to ascertain the prevalence and characteristics of MGUS among Chinese population. Methods A total of 154597 consecutive healthy participants from Beijing who underwent annual physical examination between December 2013 and April 2019 at Peking Union Medical College Hospital were enrolled. Serum M protein was evaluated by capillary electrophoresis. Patients with a positive or suspicious serum M protein were suggested to be referred to the hematological clinic for immunofixation electrophoresis (IFE) and free light chain (FLC) assays. MGUS was defined in accordance with previous definitions. We calculated age-specific and sex-specific prevalence and described laboratory characteristics of patients with MGUS among those participants. Results MGUS were diagnosed in 843 patients (0.55%, 95%CI 0.51% to 0.59%). The median age at presentation was 58 years, with a range of 25-96 years. The overall prevalence of MGUS was 1.14% among participants aged 50 years or older and 2.6% among those aged 70 years or older. In both sexes, the prevalence increased with age: 0.1% (<40 years), 0.36% (40-49 years), 0.78% (50-59 years), 1.28% (60-69 years), 2.19% (70-79 years), and 3.77% (≥80 years) separately (Figure 1). The prevalence among men were higher than that among women (0.67% vs. 0.40%, OR =1.719, 95% CI 1.490 to 1.983, P<0.001) (Figure 1). The median concentration of serum Monoclonal protein was 1.4 g/L (0.1 -27.8 g/L). M protein level was less than 0.5g/L in 220 patients (26.1%), less than 5 g/L in 81.1% and more than 15 g/L in only 1.9% of 843 persons. There was no significant difference in the concentration of the monoclonal protein among the age groups. Of the 519 patients who were tested for IFE, the isotype of the monoclonal immunoglobulin was IgG in 344 (66.3%), IgA 112 (21.6%), IgM in 48 (9.2%), IgD in 2 (0.4%), light-chain in 3 (0.6%) and biclonal in 10 (1.9%). The serum light-chain type was kappa in 260 (50.1%), lambda in 255 (49.1%) patients, while 4 patients (0.8%) with biclonal M protein have both kappa and lambda light-chain. Of the 180 people who were tested for FLC, 42 (23.3%) had an abnormal FLC ratio. IgG isotype, M protein <15 g/L and normal FLC ratio were found in 102 patients (56.7%) and the remaining 78 people (43.4%) had 1(30.6%) or 2(12.8%) abnormal factors. Conclusions MGUS was found in 1.14% of persons 50 years of age or older and 2.6% among those 70 years of age or older among healthy Chinese population. The prevalence of MGUS increases with age. Males have a higher frequency of MGUS than Females. These observations offer the overall situation of MGUS epidemiology in a large Chinese population. Disclosures No relevant conflicts of interest to declare.

Monoclonal protein reference change value as determined by gel-based serum protein electrophoresis

2018 ◽  
Vol 51 ◽  
pp. 61-65 ◽  
Author(s):  
Mina Salamatmanesh ◽  
Christopher R. McCudden ◽  
Arleigh McCurdy ◽  
Ronald A. Booth
Keyword(s):  
Serum Protein ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Monoclonal Protein ◽  
Reference Change Value

Effect of Different Integration Approaches to the M-Spike Quantification in Serum Protein Electrophoresis and How It Correlates to the New Heavy/Light Chain Assay

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5345-5345
Author(s):  
Juana Jiménez Jiménez ◽  
Tiago Pais ◽  
Luisa Campos ◽  
Carmen Hernando de Larramendi
Keyword(s):  
Binding Site ◽  
Serum Protein ◽  
Light Chain ◽  
Relative Difference ◽  
Protein Electrophoresis ◽  
Integration Methods ◽  
Monoclonal Protein ◽  
M Spike ◽  
Total Immunoglobulin ◽  
Better Than

Abstract Introduction To measure the Monoclonal Protein (MP) is essential in monoclonal gammopathies (MG) for both diagnosis and response assessment. However, the M-spike quantification may be affected by the inherent subjectivity and by the opted integration method to determine the area under the M-spike corresponding to the MP in a serum protein electrophoresis (SPE). Co-migration with other serum proteins and strong polyclonal background may also affect the MP accurate measurement. Due to its pivotal role in managing MG, and since new automatized techniques have been developed for the MP quantification, this study aims at compare different M-spike integration methods on the measurement of MP, and compare it to the new Heavy/Light chain assay (HLC/ Hevylite®), which allows to separately quantify, by automated nephelo/turbidimetry, the serum levels of IgG-K, IgG-L, IgA-K, IgA-L, IgM-K and IgM-L. Material and Methods 147 samples from 143 MGUS and MM patients were included. All the samples were analyzed by SPE (SEBIA- HYDRASYS 2) and two integration methods were used for quantifying the MP: MP1: peak defined until baseline; MP2: peak defined excluding the polyclonal part. The size of the paraprotein measured by the MP1 method was used to divide the samples in MP<10g/L and MP>10g/L. Statistical analysis included Passing-Bablok (PB) and Bland-Altman (Analyse-It®) and Mann-Whitney test (GraphPad Prism). Results The MP quantification by the MP1 and MP2 methods was significantly different (medians: 7.4 g/L vs 4.1 g/L, P<0.0001), with a relative difference of -64.36%. When dividing samples according to the MP size, the mean relative difference found was higher in samples with MP<10g/L than MP>10g/L (MP1:-85.02% vs MP2:-28.23%; medians: 5.4g/L vs 2.2g/L, P<0.0001; and 18.45g/L vs 14.65g/L, P=0.0253; respectively). Likewise, the correlation between MP1 and MP2 was good (r=0.991) but it weakened to r=0.864 in samples with MP< 10g/L. Therefore, the two integration methods diverged more in the quantification of small MP. In all analysis, MP1 correlated better than the MP2 method with the involved HLC (iHLC) (r=0.886 and r=0.87, respectively) and the total Ig (r=0.924 and r=0.9, respectively), with lower mean relative differences. The MP1 correlation with iHLC was considerably better in samples with MP>10g/L in respect to MP<10g/L (r=0.837 and r=0.55, respectively). PB analysis further revealed an agreement between MP1 and iHLC quantification in samples with MP>10g/L: iHLC=2.096+1.045PM1 (95%CI: Intercept -3.978 to 5.719; Slope: 0.8568 to 1.320), confirming that Hevylite is a useful tool for the MP quantification. Moreover, PB analysis on the summation of Heavy/Light pairs of the same immunoglobulin isotype resulted in an agreement with the levels of total Immunoglobulin (tIg) (r=0.857. PB: ∑HLC=0.1819+0.9211tIg; 95%CI: Intercept -0.9733 to 1.331; Slope: 0.8454 to 1.006), being slightly better in MP<10g/L than >10g/L, indicating the HLC analysis is valid for small MP measurements. HLC analysis also allows measuring the uninvolved-HLC pair (uHLC), a new immunoparesis parameter than has been suggested to have prognostic value. In this study, 32/34 MM and 49/109 MGUS samples had suppressed levels of uHLC. Interestingly, the MP1 and MP2 correlation with iHLC was better in MM than in GMSI samples (r=0,864 and r=0,857, versus, r=0,678 and r=0,606, respectively), which may be due both to generally higher levels of MP or lower polyclonal background in MM samples. Furthermore, 2/109 MGUS samples resulted in a different risk-of-progression category depending on the method of M-spike integration, with the iHLC being in agreement with the MP1 in one of the samples and discordant with the other. Conclusions The MP measurement by M-spike integration until baseline correlated with the iHLC and total immunoglobulin measurement better than when the M-spike integration excludes the polyclonal part. The good agreement between the iHLC and SPE in the quantification of the monoclonal protein puts this assay as an alternative method especially relevant at low concentration levels, which may help correcting for methodology associated variability within clinical laboratories. Finally, HLC may be easier to standardize than the SPE analysis and could eliminate the need for total Ig quantification. Disclosures Pais: The Binding Site Spain: Employment. Campos:The Binding Site Spain: Employment.

scholarly journals An international multi-center serum protein electrophoresis accuracy and M-protein isotyping study. Part II: limit of detection and follow-up of patients with small M-proteins

2020 ◽  
Vol 58 (4) ◽  
pp. 547-559 ◽  
Author(s):  
Joannes F.M. Jacobs ◽  
Katherine A. Turner ◽  
Maria Stella Graziani ◽  
Jody L. Frinack ◽  
Michael W. Ettore ◽  
...  
Keyword(s):  
Serum Protein ◽  
Protein Concentration ◽  
Limit Of Detection ◽  
Negative Impact ◽  
Protein Electrophoresis ◽  
Protein Quantification ◽  
Serum Protein Electrophoresis ◽  
M Protein ◽  
M Proteins

AbstractBackgroundElectrophoretic methods to detect, characterize and quantify M-proteins play an important role in the management of patients with monoclonal gammopathies (MGs). Significant uncertainty in the quantification and limit of detection (LOD) is documented when M-proteins are <10 g/L. Using spiked sera, we aimed to assess the variability in intact M-protein quantification and LOD across 16 laboratories.MethodsSera with normal, hypo- or hyper-gammaglobulinemia were spiked with daratumumab or elotuzumab, with concentrations from 0.125 to 10 g/L (n = 62) along with a beta-migrating sample (n = 9). Laboratories blindly analyzed samples according to their serum protein electrophoresis (SPEP)/isotyping standard operating procedures. LOD and intra-laboratory percent coefficient of variation (%CV) were calculated and further specified with regard to the method (gel/capillary electrophoresis [CZE]), gating strategy (perpendicular drop [PD]/tangent skimming [TS]), isotyping (immunofixation/immunosubtraction [ISUB]) and manufacturer (Helena/Sebia).ResultsAll M-proteins ≥1 g/L were detected by SPEP. With isotyping the LOD was moderately more sensitive than with SPEP. The intensity of polyclonal background had the biggest negative impact on LOD. Independent of the method used, the intra-laboratory imprecision of M-protein quantification was small (mean CV = 5.0%). Low M-protein concentration and high polyclonal background had the strongest negative impact on intra-laboratory precision. All laboratories were able to follow trend of M-protein concentrations down to 1 g/L.ConclusionsIn this study, we describe a large variation in the reported LOD for both SPEP and isotyping; overall LOD is most affected by the polyclonal immunoglobulin background. Satisfactory intra-laboratory precision was demonstrated. This indicates that the quantification of small M-proteins to monitor patients over time is appropriate, when subsequent testing is performed within the same laboratory.

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