What is SPEP (Serum Protein Electrophoresis) / UPEP (Urine Protein Electrophoresis)?

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Last updated: December 11, 2025View editorial policy

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What is SPEP/UPEP?

SPEP (Serum Protein Electrophoresis) and UPEP (Urine Protein Electrophoresis) are laboratory tests that separate and measure different proteins in blood and urine to detect and quantify monoclonal proteins (M-proteins), which are abnormal antibodies produced by clonal plasma cells in disorders like multiple myeloma. 1

How These Tests Work

SPEP separates serum proteins based on their physical properties and electrical charge, creating distinct bands representing different protein fractions (albumin, alpha-1, alpha-2, beta, and gamma globulins). 2 When a monoclonal protein is present, it appears as a sharp, homogeneous spike (M-spike) typically in the gamma region, though it can appear in the beta region. 1, 3

UPEP performs the same separation technique on a 24-hour urine collection to detect M-proteins that are excreted in urine, particularly light chains (Bence Jones proteins). 4

Primary Clinical Applications

These tests serve four main purposes in plasma cell disorder evaluation:

  • Initial screening for suspected multiple myeloma, Waldenström's macroglobulinemia, monoclonal gammopathy of undetermined significance (MGUS), and related conditions 1
  • Quantifying M-protein levels to establish baseline disease burden 4
  • Monitoring treatment response by tracking changes in M-protein concentration over time 4
  • Detecting disease recurrence during surveillance 1

Diagnostic Workflow After Abnormal Results

When SPEP shows an M-spike, the National Comprehensive Cancer Network mandates specific follow-up testing: 1

  • Serum immunofixation electrophoresis (SIFE) to identify the exact immunoglobulin type (IgG, IgA, IgM) and light chain (kappa or lambda) 4, 1
  • Quantitative immunoglobulin levels (IgG, IgA, IgM) to assess immune paresis 1
  • Serum free light chain (FLC) assay with kappa/lambda ratio 4, 1
  • 24-hour urine collection for total protein, UPEP, and urine immunofixation electrophoresis (UIFE) 4, 1

Critical Limitations and Pitfalls

SPEP has significant sensitivity limitations. In patients with lytic bone lesions, SPEP demonstrates only 71% sensitivity and 47% positive predictive value for plasma cell neoplasms, though it maintains 94% negative predictive value. 5 This means a negative SPEP does not rule out myeloma.

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 An additional 3% of patients have truly non-secretory myeloma with no detectable M-protein on either SPEP or UPEP. 4

The combination of SPEP plus serum free light chain analysis achieves 100% sensitivity for detecting plasma cell disorders, superior to SPEP plus UPEP. 6 This modern approach can eliminate the need for 24-hour urine collection in initial screening, though UPEP remains necessary for monitoring patients with measurable urinary M-proteins. 4

Interpretation Nuances

Renal impairment causes false-positive elevations in serum free light chain ratios up to 3.1 (normal 0.26-1.65) without true plasma cell disorder, requiring interpretation in context of serum creatinine levels. 7

Polyclonal gammopathies (broad-based elevation in gamma region without sharp spike) indicate reactive or inflammatory processes, not clonal plasma cell disorders, and must be distinguished from monoclonal patterns. 2

When to Refer

Any detected monoclonal protein on SPEP requires hematology/oncology referral. 1 Urgent referral within 1-2 weeks is mandatory for patients with significant M-protein spike accompanied by anemia, renal dysfunction, hypercalcemia, or bone lesions. 1

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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