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Interferencia y fallo: El desafío de la proteína monoclonal IgM de alta concentración en la electroforesis capilar de

Yanhua Zhao¹, Minjie Zhang¹, Wei Gan¹

¹Department of Laboratory Medicine, West China Hospital, Sichuan University, Sichuan Clinical Research Center for Laboratory Medicine, and Clinical Laboratory Medicine Research Center of West China Hospital, Chengdu 610041, Sichuan Province, China.

Clinical biochemistry

|February 17, 2026

Insights

Los niveles altos de IgM en la macroglobulinemia de Waldenström pueden causar fallos en la electroforesis capilar de zona (CZE). El pretratamiento con beta-mercaptoetanol (BME) resolvió con éxito la interferencia, previniendo resultados falsos negativos y asegurando un diagnóstico preciso.

Área de la Ciencia:

Química Clínica
Electroforesis de Proteínas
Inmunología

Sus antecedentes:

La electroforesis capilar de zona (CZE) es crucial para la separación de proteínas séricas.
Las proteínas monoclonales IgM pueden interferir con el CZE, lo que lleva a resultados inexactos.
La macroglobulinemia de Waldenström presenta un desafío diagnóstico debido a la posible interferencia del CZE.

Objetivo del estudio:

Investigar la interferencia de altas concentraciones de IgM en el CZE.
Evaluar métodos de pretratamiento para resolver fallos de CZE en la macroglobulinemia de Waldenström.
Identificar estrategias efectivas para la detección precisa de proteínas monoclonales IgM.

Principales métodos:

Se estudió un caso de macroglobulinemia de Waldenström con fallos recurrentes de CZE.
Se evaluaron cuatro protocolos de pretratamiento: calor, dilución acuosa, dilución salina y beta-mercaptoetanol (BME).
Se utilizaron EFS en gel e CZE por inmunosustracción para analizar los mecanismos de interferencia.

Principales resultados:

El CZE estándar falló debido a altos niveles de IgM.
Los métodos de calor y dilución no resolvieron la interferencia.
El pretratamiento con BME restauró con éxito el análisis de CZE, revelando la proteína monoclonal IgM.
La EFS en gel confirmó la proteína M, mientras que el CZE por inmunosustracción indicó interferencia por crioglobulina.

Conclusiones:

Altas concentraciones de IgM, especialmente con actividad de crioglobulina, pueden causar fallos en el CZE.
La interferencia probablemente se deba a la agregación de proteínas o a la precipitación dependiente de la temperatura.
El pretratamiento con BME es esencial para el análisis preciso de CZE de proteínas monoclonales IgM, previniendo errores diagnósticos.

Read full analysis

Abstracto

Background:

Capillary zone electrophoresis (CZE) is a standard method for serum protein separation, but its performance can be compromised by IgM monoclonal proteins, leading to false-negative results.

Methods:

We report a case of Waldenström macroglobulinemia (IgM 56.05 g/L) with repeated CZE failure. Immunofixation electrophoresis (IFE) confirmed an IgM kappa monoclonal protein. Four pretreatment protocols were evaluated: 1) 37°C incubation; 2) dilution with distilled water; 3) dilution with 0.9% saline; and 4) pretreatment with β‑mercaptoethanol (BME). Gel-based serum protein electrophoresis (SPE) and immunosubtraction CZE were also performed to investigate the interference mechanism.

Results:

Initial CZE failed to yield an interpretable electrophoretogram. Incubation at 37°C showed no improvement. Dilution with distilled water induced visible precipitation and resulted in a false-negative electrophoretic pattern with a marked drop in measured total protein. Dilution with saline prevented precipitation but still failed to reveal a distinct monoclonal spike, despite biochemical measurements consistent with high globulin levels. In contrast, pretreatment with BME successfully restored a clear monoclonal spike in the β‑γ region on CZE, demonstrating restored solubility and proper migration. IFE corroborated the presence of the IgM kappa monoclonal protein. Gel-based SPE successfully showed a distinct M-protein band without interference, while immunosubtraction CZE failed under both temperature conditions, supporting cryoglobulin-related interference.

Conclusions:

Very high IgM concentrations, particularly with cryoglobulin activity, can cause complete CZE failure not resolved by standard dilution or heating. The interference likely involves capillary obstruction from protein aggregation or temperature-dependent re-precipitation within the capillary. BME pretreatment is a critical intervention for unmasking IgM monoclonal proteins when CZE results are inconsistent with other laboratory findings, thereby preventing diagnostic errors.

Palabras clave:

Electroforesis capilar de zona Crioglobulina Falso negativo Gammopatía monoclonal IgM β‑mercaptoetanol

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