What is Serum Protein Electrophoresis (SPEP) testing?

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What is SPEP Testing?

Serum Protein Electrophoresis (SPEP) is a laboratory test that separates blood proteins based on their electrical charge and size to detect and quantify monoclonal proteins (M-proteins), primarily used to diagnose and monitor plasma cell disorders such as multiple myeloma, as well as evaluate polyneuropathies and other protein abnormalities. 1

Primary Clinical Applications

Detection of Monoclonal Gammopathies

  • SPEP identifies the presence, type, and quantity of M-proteins (abnormal antibodies produced by clonal plasma cells), appearing as a characteristic monoclonal spike (M-spike) on the electrophoresis pattern, typically in the gamma region. 1
  • The test is fundamental for diagnosing multiple myeloma, Waldenström's macroglobulinemia, monoclonal gammopathy of undetermined significance (MGUS), and related plasma cell disorders. 1, 2
  • Monoclonal gammopathies are significantly more common in patients with polyneuropathy (10% prevalence) compared to the general population, making SPEP a high-yield screening test in this context. 3

Evaluation of Polyneuropathy

  • SPEP is one of the three highest-yield screening tests for distal symmetric polyneuropathy (DSP), alongside blood glucose and serum B12 with metabolites. 3
  • However, serum protein immunofixation electrophoresis (IFE) is more sensitive than SPEP, especially for detecting small or non-malignant monoclonal gammopathies—IFE identified 17% of monoclonal gammopathies that SPEP missed. 3

How the Test Works

Technical Methodology

  • SPEP separates serum proteins into distinct fractions based on their physical properties: albumin, alpha-1 globulin, alpha-2 globulin, beta globulin, and gamma globulin. 2, 4
  • The resulting pattern displays reasonably predictable changes in response to acute inflammation, malignancy, trauma, necrosis, infarction, burns, and chemical injury. 2
  • A homogeneous spike-like peak in a focal region (usually gamma-globulin zone) indicates a monoclonal gammopathy requiring further investigation. 2

Interpretation and Follow-Up

Abnormal Findings

  • When SPEP shows an M-spike or abnormal pattern, mandatory follow-up testing includes serum immunofixation electrophoresis (SIFE) to identify the exact immunoglobulin type (IgG, IgA, IgM) and light chain (kappa or lambda). 1
  • Additional required tests include quantitative immunoglobulin levels, serum free light chain (FLC) assay with kappa/lambda ratio, and 24-hour urine collection for urine protein electrophoresis (UPEP) and urine immunofixation. 1
  • The combination of SPEP plus serum free light chain analysis achieves 100% sensitivity for detecting plasma cell disorders. 1

Clinical Significance of M-Protein Levels

  • MGUS typically shows M-protein less than 30 g/L, while multiple myeloma typically shows higher levels. 1
  • The height and area under the M-spike can be quantified to track disease burden and monitor treatment response. 1

Important Limitations and Pitfalls

Sensitivity Concerns

  • Approximately 15-20% of myeloma cases produce only light chains (not complete antibodies), which may not create a visible spike on standard SPEP and require urine testing or serum free light chain assays for detection. 1
  • About 3% of patients have truly non-secretory myeloma with no detectable M-protein on either SPEP or UPEP, highlighting the importance of additional diagnostic tests. 1
  • Serum immunofixation electrophoresis is more sensitive than SPEP and should be used when clinical suspicion is high despite negative SPEP. 3

Referral Indications

  • Any detected monoclonal protein on SPEP should prompt referral to a hematologist/oncologist, with urgent referral (within 1-2 weeks) recommended for patients with significant M-protein spike, accompanying symptoms, anemia, renal dysfunction, or hypercalcemia. 1

Monitoring Applications

Disease Surveillance

  • SPEP is used to quantify M-protein levels at baseline to establish disease burden. 1
  • Serial SPEP measurements track changes in M-protein concentration over time to monitor treatment response. 1
  • The test detects disease recurrence during surveillance in patients with known plasma cell disorders. 1

References

Guideline

Serum Protein Electrophoresis in Plasma Cell Disorders

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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