Evusheld, a SARS-CoV-2 spike protein-directed attachment inhibitor, appears in serum protein electrophoresis and immunofixation: a case study.

Serum protein electrophoresis (SPE) and immunofixation (IFE) assays are commonly used to diagnose and monitor patients with multiple myeloma (MM). Identifying analytical interferences in SPE and IFE caused by therapeutic monoclonal antibodies (tmAbs) can be challenging. Here we report the case of a 72-year-old male with a long history of relapsed immunoglobulin (Ig)G kappa MM. A follow-up SPE showed the original peak plus 2 additional cathode peaks. Immunofixation was ordered as a reflex test to investigate the new peaks that showed initial patient monoclonal IgG kappa in addition to 2 restricted bands of the IgG kappa type. Therapeutic monoclonal antibody interference was suspected and the patient's chart was reviewed. The patient was not on any antimyeloma monoclonal antibody therapy. However, preexposure prophylaxis therapeutic monoclonal antibodies tixagevimab plus cilgavimab (Evusheld) for severe acute SARS-CoV-2 was administered approximately 45 minutes before sample collection, which led to the identifiable spikes and correlated bands. After 2 days, the IgG kappa bands disappeared, confirming this therapy's effect on SPE and IFE. Therefore, clinical pathologists should be aware of when providers prescribe new monoclonal antibody therapy and become familiar with the position of commonly prescribed (tmAbs) therapies at their institutions.

[Detection of paraprotein in plasma cell tumors].

Paraprotein is a laboratory biomarker of plasma cell tumors and other lymphoproliferative diseases. Its determination is necessary for diagnosing, monitoring and assessment of therapy effectiveness. The lecture presents the main methods of qualitative and quantative analysis of monoclonal proteins: gel electrophoresis, capillary electrophoresis, immunofixation and nephelometry features, possibilities and limitations are reviewed. The main sources of errors and artifacts during these studies are considered. Also the difficulties in the diagnosis and interpretation of the results of serum and urine tests are highlighted.

To skim or splice? Comparing the quantification of M-proteins using two peak-integration protocols across multiple electrophoresis platforms.

OBJECTIVES: M-protein quantification by peak integration in serum protein electrophoresis (SPE) plays a central role in diagnosing, prognosing and monitoring monoclonal gammopathies. The conventional perpendicular drop (PD) integration approach integrates M-spikes from the baseline, which performs acceptably when the M-protein concentration is relatively high compared to the amount of background proteins present. The alternative peak-integration protocol by tangential skim (TS), however, allows for more accurate M-protein estimations by excluding background proteins. Despite some guideline recommendations, TS has been poorly adopted, making an understanding of the differences between the two protocols and their potential impacts paramount when considering a change from PD to TS. DESIGN & METHODS: We conducted retrospective investigations of the differences in M-protein quantification over large concentration ranges between PD and TS on 3 of the most popular electrophoresis platforms. RESULTS: Compared to PD, TS gave consistently lower results; the differences between the two methods increased tremendously and became more sporadic as M-protein concentrations dropped below 15 g/L in all 3 platforms. At < 15 g/L, the average % difference ranged from -81 % to -95 %, while above 15 g/L, the average % difference was only -13 to -31 %. Medical decision point analyses using linear regression predicted statistically significant and platform-dependent differences, which could impact clinical interpretation. CONCLUSIONS: Careful consideration of the magnitude of concentration changes and the potential impacts on patient classification and management should be made when switching to TS for M-protein quantification.

Detection of Plasma Cell Disorders by Mass Spectrometry: A Comprehensive Review of 19,523 Cases.

OBJECTIVES: To verify the analytical performance of a new mass spectrometry-based method, termed MASS-FIX, when screening for plasma cell disorders in a routine clinical laboratory. PATIENTS AND METHODS: Results from 19,523 unique patients tested for an M-protein between July 24, 2018, and March 6, 2020, by a combination serum protein electrophoresis (SPEP) and MASS-FIX were examined for consistency with pretest implementation performance. MASS-FIX's ability to verify abnormal results from SPEP and free light chain measurements was then compared with that of immunofixation electrophoresis (IFE) using a separate cohort of 52,586 patients tested by SPEP/IFE during the same period. RESULTS: Overall, 62.4% of our cohort was negative for an M-protein. Importantly, 7.3% of all specimens had an M spike on SPEP (0.1 to 8.5 g/dL) and MASS-FIX detected an M-protein in all these samples. Of all samples, 30.3% had M-proteins that were detected by MASS-FIX but the SPEP finding was too small for quantification. Of the positive samples, 5.7% contained a therapeutic monoclonal antibody. Of the positive samples, 4.1% had an N-glycosylated light chain (biomarker of high-risk plasma cell disorders). MASS-FIX confirmed a higher percentage of SPEP abnormalities than IFE. MASS-FIX was slightly more sensitive than IFE when confirming an M-protein in samples with an abnormal free light chain ratio. MASS-FIX had a very low sample repeat rate (1.5%). MASS-FIX was highly automatable resulting in a higher number of samples/technologist/day than IFE (∼30% more). CONCLUSION: Overall, MASS-FIX was successful in maintaining validation characteristics. MASS-FIX was more sensitive in confirming SPEP abnormalities when compared with IFE. Ability to detect therapeutic monoclonal antibodies and glycosylated light chains was distinctly advantageous.

Accurate Quantification of Monoclonal Immunoglobulins Migrating in the Beta Region on Protein Electrophoresis.

BACKGROUND: The concentration of monoclonal immunoglobulins (Igs) in neoplastic monoclonal gammopathic manifestations is generally measured by densitometric scanning of the monoclonal peaks on gel or by measuring absorbance at 210 nm in capillary electrophoresis (CE). For monoclonal Igs migrating in the beta region, measurement is complicated by the major beta-region proteins, namely, transferrin and C3. METHODS: C3 interference in densitometry was eliminated by heat treatment of serum, and monoclonal Igs were quantified by densitometry of the residual band. The immunochemical measurement of transferrin was converted to its equivalent densitometric quantity. For monoclonal Ig migrating with transferrin, the contribution of the latter was removed by subtracting the converted transferrin concentration from the combined densitometric quantification of the band. With CE, monoclonal Ig was measured by using immunosubtraction (ISUB) to guide demarcation. RESULTS: The results obtained using the C3 depletion and transferrin subtraction method were lower and yet comparable to the results derived from using CE measurement guided by ISUB. As we expected, the results from both methods were lower than those derived from a perpendicular drop measurement of the peak or via nephelometric assay of the involved isotype. DISCUSSION: Accurate measurement of monoclonal Igs is important for the diagnosis and monitoring of monoclonal gammopathic manifestations. Determination of serum free light chain concentration per gram of monoclonal Ig is an essential measure for the diagnosis of light chain-predominant multiple myeloma. The method described herein improves accuracy of measurements for monoclonal Igs migrating in the beta region, without the need for special reagents or equipment.

Application of an optimized interpretation model in capillary hemoglobin electrophoresis for newborn thalassemia screening.

INTRODUCTION: Newborn screening is an important supplement to thalassemia control and prevention. Capillary electrophoresis (CE) technology has several advantages for thalassemia screening but with low sensitivity, especially for thalassemia carriers. This study aims to illustrate the application of an optimized interpretation model in newborn thalassemia screening by capillary hemoglobin electrophoresis. METHODS: Two thousand, two hundred fifty-eight neonates selected from four regions in China were enrolled and were screened for α-thalassemia and ß-thalassemia by capillary electrophoresis. Results were interpreted based on an optimized model integrated with multiple parameters. Molecular analysis was carried out in synchrony and used as the gold standard for the screening performance assessment. The consistency among different regions and thalassemia genotypes were also investigated. RESULTS: Among the 2258 neonates, 485 were identified to have a likely diagnosis of thalassemia, and 422 α-thalassemia, 80 ß-thalassemia, and 21 α/ß-thalassemia cases were confirmed by molecular analysis, including 277 α-thalassemia silent carriers, 135 α-thalassemia trait carriers, 10 Hemoglobin H disease, and 80 ß-thalassemia trait carriers. The screening sensitivity, specificity, positive, and negative predictive value for α-thalassemia and ß-thalassemia were 84.83%, 99.14%, 95.98%, 96.41%, and 88.75%, 98.73%, 76.34%, and 99.48%, respectively. The optimized interpretation model showed higher performance for thalassemia carriers, though some neonates with silent α-thalassemia genotypes (-α3.7 /αα, -α4.2 /αα, and αWS α/αα) and ß-28 /ßN genotype were still missed. The screening performance among different regions was comparable. CONCLUSIONS: Capillary hemoglobin electrophoresis with the optimized interpretation model shows reliable performance for newborn thalassemia screening. It is applicable to large-scale population screening.

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

OBJECTIVES: Monoclonal immunoglobulins (M-proteins) that migrate in the ß 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 ß-region bands for detection of ß-migrating M-proteins. METHODS: Consecutive SPEP cases analyzed by capillary electrophoresis were searched to identify the initial workup on 1,841 patients with increased total ß regions, suspicious ß-region findings resulting in reflex immunofixation (IFE), or immunosubtraction (ISUB). To augment quantitative information, separate ß1 and ß2 measurements were established and retrospectively used to evaluate their sensitivity for M-protein detection. RESULTS: 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 ß1 and ß2 measurements that were not identified by total ß-region measurement, 1 progressed to myeloma. Of the 56 ß-migrating M-proteins identified by qualitative features but without increase in any of the ß-region measurements, 1 progressed to myeloma. CONCLUSIONS: A combination of separate measurements for ß1 and ß2 regions together with detection of ß-region distortions increase sensitivity for identifying ß-migrating M-proteins via reflex IFE or ISUB.

Estudio comparativo: cromatografía liquida de alta presión versus electroforesis de hemoglobina. Diagnóstico de anemia drepanocítica / Comparative study: high pressure liquid chromatography versus hemoglobin electrophoresis. Diagnosis of sickle cell anemia

La anemia de células falciformes (ACF) es la hemoglobinopatía hereditaria más común en el mundo. El diagnóstico definitivo se hace por electroforesis de hemoglobina (EFH), relativamente costosa. Por ello ante la sospecha diagnóstica se solicita Metabisulfito de sodio al 2% (MS2), que tiene un menor costo, aporta solo resultados cualitativos, y la confiabilidad depende de quien interprete. Se busca una alternativa económicamente accesible y con mayor certeza diagnóstica. Objetivo: describir y comparar los valores de Hemoglobina S (HbS) obte- nidos mediante cromatografía líquida de alta presión (CLAP) y electroforesis de hemoglobina. Pacientes y Métodos: investigación cuantitativa, descriptiva, no experimental, en las comuni- dades de Masca y Pueblo Nuevo, Omoa, Cortés, en el 2017. Las comunidades tienen 2545 habitantes, de quienes se tomó muestra probabilística de 369. Encontrando 20 personas con prueba de MS2 positiva. En estos 20 casos se solicitó también CLAP y EFH. Los datos fueron analizados con SPSS versión 23, calculando frecuencias, porcentajes y medidas de tendencia central. Resultados: El 50% de los casos eran fenotípicamente mestizos. Los valores de CLAP estuvieron comprendidos entre 26.1% y 68.3% y los de electroforesis de hemoglobina entre 27.3% y 100%. La media aritmética de CLAP fue de 35.53 vs 45.3 para EFH. Conclu- sión: El valor de Hb S medido por EFH es cercano al obtenido por CLAP, por lo que este método podría usarse para un diagnóstico más rápido y a menor costo...(AU)

Quantification of Complement Proteins with Special Reference to C1q: Multiplex Versus ELISA Versus Rocket Immunoelectrophoresis Versus Nephelometry.

Accurate determination of complement component C1q is hampered by the fact that C1q is an immune complex binding protein. Consequently, immunochemical techniques which rely on immune complex formation in fluid phase such as nephelometry and turbidimetry tend to give results which differ from those obtained by, for example, ELISA and other solid phase-based assays. In this chapter, we discuss the pros and cons of different techniques for the quantification of C1q and present a comprehensive protocol for a newly developed magnetic bead-based sandwich immunoassay which has replaced nephelometry in our complement diagnostic laboratory at the University Hospital in Uppsala.

OCCURRENCE OF UNUSUAL HAEMOGLOBINOPATHIES IN BALOCHISTAN: HB SD AND HB SE - PRESENTATION WITH OSTEOMYELITIS.

OBJECTIVE: To describe two cases of unusual variants of sickle cell disease. CASE DESCRIPTION: We present two cases of sickle cell disease variants (haemoglobinopathies), from unrelated families, in the state of Balochistan (Pakistan). One was diagnosed with sickle cell disease in the haemoglobin electrophoresis, whereas the other was diagnosed with sickle cell SE disease. Both were diagnosed based on the presentation of osteomyelitis. COMMENTS: Haemoglobin SD disease (Hb SD) and haemoglobin SE disease (Hb SE) are rare haemoglobinopathies in the world. The lack of available literature suggests that both are variants of sickle cell disease (SCD), with heterogeneous nature. The prevalence of sickle cell disease with compound heterozygotes was found at a variable frequency in the population of the Asian Southeast. The frequency of osteomyelitis in SCD is 12 to 18%, but its occurrence among variant haemoglobinopathies is little reported. Both reported cases presented with osteomyelitis as a characteristic of the disease presentation.

Baseline correlations and prognostic impact of serum monoclonal proteins in follicular lymphoma.

The presence of a serum monoclonal component has been associated with poor outcomes in some lymphomas. However, data in follicular lymphoma (FL) are scarce. We studied 311 FL patients diagnosed at a single institution, for whom information on serum immunofixation electrophoresis (sIFE) at diagnosis was available. Baseline characteristics and outcomes were compared between patients with a positive (+sIFE) and a negative sIFE (-sIFE). sIFE was positive in 82 patients (26%). Baseline features were comparable between both groups, except for an older age and higher proportion of elevated ß2 -microglobulin levels in the +sIFE group. With a median follow-up of 4.6 years, a +sIFE was associated with a higher risk of early relapse (POD24, 27% vs. 15%, P = 0·02), shorter progression-free survival (PFS; 42% vs. 52% at 5 years, P = 0·008), and shorter overall survival (OS; 59% vs. 77% at 10 years, P = 0·046). In patients >60 years, a +sIFE was an independent predictor of OS [hazard ratio (HR) = 2·4, 95% confidence interval (CI): 1·2-5·0; P = 0·02]. Approximately one quarter of patients with FL has a +sIFE at diagnosis, which is a predictor of poor outcome. These findings encourage further investigation of its relationship with B-cell biology and the tumour microenvironment.

Occurrence of unusual haemoglobinopathies in balochistan: hb sd and hb se - presentation with osteomyelitis / Ocorrência de hemoglobinopatias incomuns no baluchistão: hb sd e hb se - apresentação com osteomielite

ABSTRACT Objective: To describe two cases of unusual variants of sickle cell disease. Case description: We present two cases of sickle cell disease variants (haemoglobinopathies), from unrelated families, in the state of Balochistan (Pakistan). One was diagnosed with sickle cell disease in the haemoglobin electrophoresis, whereas the other was diagnosed with sickle cell SE disease. Both were diagnosed based on the presentation of osteomyelitis. Comments: Haemoglobin SD disease (Hb SD) and haemoglobin SE disease (Hb SE) are rare haemoglobinopathies in the world. The lack of available literature suggests that both are variants of sickle cell disease (SCD), with heterogeneous nature. The prevalence of sickle cell disease with compound heterozygotes was found at a variable frequency in the population of the Asian Southeast. The frequency of osteomyelitis in SCD is 12 to 18%, but its occurrence among variant haemoglobinopathies is little reported. Both reported cases presented with osteomyelitis as a characteristic of the disease presentation.

RESUMO Objetivo: Descrever dois casos de variantes raras da hemoglobinopatia falciforme. Descrição do caso: Apresentamos aqui dois casos de hemoglobinopatias variantes das células falciformes, de famílias não relacionadas, no estado do Baluchistão (Paquistão), sendo um diagnosticado como doença da hemoglobina SD na eletroforese de hemoglobina, enquanto o outro com doença da hemoglobina SE. Ambos foram diagnosticados a partir da apresentação de osteomielite. Comentários: Hemoglobina SD (Hb SD) e hemoglobina SE (Hb SE) são hemoglobinopatias raras no mundo. A escassez de literatura disponível sugere que ambas são variantes da doença falciforme (DF) com natureza heterogênea. A prevalência de hemoglobinopatia falciforme com heterozigosidade composta foi encontrada com frequência variável na população do sudeste asiático. A frequência de osteomielite na DF é de 12 a 18%, mas sua ocorrência entre as hemoglobinopatias falciformes variantes é pouco relatada. Os dois casos reportados apresentaram osteomielite como característica de apresentação da doença.

Enhancing the Role of the Medical Technologist in Serum Protein Electrophoresis Interpretation: A Pattern Recognition Approach to Identifying Obvious and Potentially Significant Subtle Alterations.

Medical technologists who perform serum protein electrophoresis can provide valuable input and assistance in the interpretation of the electrophoretic pattern. Recognition of common alterations in conjunction with the utilization of standardized interpretation comments expedites the final interpretation performed by the clinical pathologist. Improved understanding of the clinical utility of the electrophoretic interpretation enhances the role and satisfaction of the medical technologists.

Una visión inmunológica del déficit de alfa 1 antitripsina / No disponible

No disponible

Measurement of Monoclonal Immunoglobulin Protein Concentration in Serum Protein Electrophoresis: Comparison of Automated vs Manual/Human Readings.

BACKGROUND: Protein concentration of monoclonal immunoglobulin in plasma-cell myeloma/multiple myeloma provides an estimate of the tumor mass and allows for monitoring of the response to treatment. Accurate and reproducible estimates of the monoclonal immunoglobulin concentration are important for patient care. OBJECTIVE: To address the optimum method for estimation of the concentration of monoclonal immunoglobulins. METHODS: Serum protein electrophoresis and immunofixation electrophoresis were conducted by using the Helena SPIFE Touch instrument. Estimation of the protein concentration of monoclonal immunoglobulin in the gamma region by computer-assisted reading was compared with the reading by technologists and pathology residents, in 300 gels. The data were compared using t-testing and analysis of variance. RESULTS: Computer-generated readings had a consistent positive bias. The correlation coefficient of the average reading by technologists and residents with the computer generated value was 0.997. The average positive bias by the computer reading was 0.29 g per dL. The intercept on the regression analysis was 0.22 g per dL. The reading by the computer was significantly higher than each of the human-interpreted readings. The readings by the 3 human groups were not significantly different amongst them. The main reason for the higher reading by the computer was inclusion of a greater area on the anodal size of the peak on the densitometric scan. CONCLUSIONS: Human- and computer-interpreted readings of the protein concentration of monoclonal immunoglobulin have a high degree of correlation. The consistent positive bias by the computer reading occurred due to inclusion of a greater area of the densitometric scan on the anodal side of the peak. We suggest that vendors should adjust such computer programs to provide readings comparable to those generated by expert humans. We recommend manual delineation of the monoclonal peaks for measuring the concentration of monoclonal immunoglobulins.

Serum Free Light Chain Assay: Shift Toward a Higher κ/λ Ratio.

BACKGROUND: The analysis of serum free light chains (FLCs) is clinically relevant for the diagnosis and therapeutic management of clonal plasma cell disorders. This study compares the performance of monoclonal and polyclonal FLC κ and λ assays in clinical samples determined in a single academic center. METHODS: Serum FLCs were analyzed from 102 patients using the Freelite (Binding Site) and N Latex (Siemens) assays on the BN ProSpec System (Siemens). When available, data for protein electrophoresis, immunofixation, C-reactive protein, and estimated glomerular filtration rate (eGFR) were combined with FLC results to evaluate performance. RESULTS: Method evaluation showed acceptable imprecision and inaccuracy measures of <4.4% and 12.9%, respectively. Poor agreement between the methods was observed, including constant and proportional bias and poor correlation (Kendall τ, 0.671-0.901). The N Latex assay was not affected by the renal impairment estimated by eGFR, unlike the FLC κ/λ ratio results by the Freelite assay. With the Freelite assay, 98% of putative controls without monoclonal gammopathy (n = 42) showed a κ/λ ratio that was above the median of the standard diagnostic range or renal diagnostic range. A shift toward higher κ/λ ratios was also observed when retrospective data between 2011 and 2017 were compared. CONCLUSIONS: Unlike the Freelite assay, κ/λ ratios analyzed with the N Latex assay were not affected by renal failure. Both methods showed acceptable performances using nephelometry, but they were poorly correlated. A shift toward κ/λ ratios might impair the specificity of borderline increased κ/λ results. This should be considered when interpreting FLC κ and λ results.

Incidence and Management of Therapeutic Monoclonal Antibody Interference in Monoclonal Gammopathy Monitoring.

BACKGROUND: The treatment of multiple myeloma (MM) has been revolutionized by the introduction of therapeutic monoclonal antibodies (tmAbs). Daratumumab, a human IgG1/κ tmAb against CD38 on plasma cells, has improved overall survival in refractory MM and was recently approved as a frontline therapy for MM. Work on tmAb interference with serum protein electrophoresis (SPE) during MM monitoring has failed to provide information for laboratories on incidence of interference and effective methods of managing the interference at a practicable level. We aimed to evaluate daratumumab and elotuzumab interference in a large academic hospital setting and implement immediate solutions. METHODS: We identified and chart reviewed all cases of possible daratumumab interference by electrophoretic pattern (120 of 1317 total cases over 3 months). We retrospectively reviewed SPE cases in our laboratory to assess clinical implications of tmAb interference before the laboratory was aware of tmAb treatment. We supplemented samples with daratumumab and elotuzumab to determine the limits of detection and run free light chain analysis. RESULTS: Approximately 9% (120 of 1317) of tested cases have an SPE and/or immunofixation electrophoresis (IFE) pattern consistent with daratumumab, but only approximately 47% (56) of these cases were associated with daratumumab therapy. Presence of daratumumab led to physician misinterpretation of SPE/IFE results. Limits of daratumumab detection varied with total serum gammaglobulin concentrations, but serum free light chain analysis was unaffected. CONCLUSIONS: Clinical laboratories currently rely on interference identification by electrophoretic pattern, which may be insufficient and is inefficient. Critical tools in preventing misinterpretation efficiently include physician education, pharmacy notifications, separate order codes, and interpretive comments.

COMPARISON OF AGAROSE GEL AND CAPILLARY ZONE ELECTROPHORESIS METHODS USING PLASMA FROM GREEN TURTLES (CHELONIA MYDAS).

Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (Chelonia mydas). Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the γ globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0-4.9% vs 2.0-28.3%) and a lower or overlapping CV in interassay tests (1.0-10.6 vs 2.3-22.0). The prealbumin, α2 globulin, and ß globulin fractions correlated the least between the methods (for all three fractions: rs ≤ 0.28, P > 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin (rs = 0.78, P < 0.0001) and γ globulins (rs = 0.78, P < 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles.

Performance comparison of EasyFix G26 and HYDRASYS 2 SCAN for the detection of serum monoclonal proteins.

BACKGROUND: Serum protein electrophoresis (SPE) is a widely used laboratory technique to diagnose patients with multiple myeloma (MM) and other disorders related to serum protein. In patients with MM, abnormal monoclonal protein can be detected by SPE and further characterized using immunofixation electrophoresis (IFE). There are several semi-automated agarose gel-based systems available commercially for SPE and IFE. In this study, we sought to evaluate the analytical performance of fully automated EasyFix G26 (EFG26) and semi-automated HYDRASYS 2 SCAN (H2SCAN) for both SPE and IFE. METHODS: Both instruments were operated according to manufacturer's instructions. Samples used include a commercially available normal control serum (NCS) and patients' specimens. The following were evaluated: precision and comparison studies for SPE, and reproducibility and comparison studies for IFE. Statistical analyses were performed using Microsoft Excel. RESULTS: For SPE repeatability study, our results showed that EFG26 has higher coefficient of variation (%CV) compared with H2SCAN for both samples except for monoclonal component with %CV of 0.97% and 1.18%, respectively. Similar results were obtained for SPE reproducibility study except for alpha-1 (4.16%) and beta (3.13%) fractions for NCS, and beta fractions (5.36%) for monoclonal sample. Subsequently, reproducibility for IFE was 100% for both instruments. Values for correlation coefficients between both instruments ranged from 0.91 to 0.98 for the five classic bands. CONCLUSION: Both instruments demonstrated good analytical performance characterized by high precision, reproducibility and correlation.