Reference Range SPE

Serum protein electrophoresis (SPEP) is an easy, inexpensive method of separating proteins

based on their net charge, size, and shape. The 2 major types of protein present in the serum are
albumin and the globulin proteins. Albumin is the major protein component of serum and
represents the largest peak that lies closest to the positive electrode.[1] Globulins comprise a much
smaller fraction of the total serum protein but represent the primary focus of interpretation of
serum protein electrophoresis. Five globulin categories are represented: alpha-1, alpha-2, beta-1,
beta-2, and gamma, with the gamma fraction being closest to the negative electrode.

The reference range for serum protein electrophoresis is as follows:

 Albumin: 3.3-5.7 g/dL

 a- 1: 0.1-0.4 g/dL

 a- 2: 0.3-0.9 g/dL

 b- 2: 0.7-1.5 g/dL

 gamma: 0.5-1.4 g/dL

The reference range may vary depending on the laboratory performing the test.

Interpretation
Various disease states or conditions alter the pattern of proteins in electrophoresis (see Table 1
below).

Table 1. Serum Protein Fractions and Conditions Associated with an Increased or Decreased
Level (Open Table in a new window)

Serum
Protein Increased Decreased
Fraction
Malnutrition, cachexia, liver disease,
Albumin Severe dehydration nephrotic syndrome, protein-losing
enteropathies, severe burns
Alpha-1 Inflammatory states, pregnancy Alpha-1 antitrypsin deficiency
Inflammatory states, nephrotic
Alpha-2 syndrome, oral contraceptive use, Hemolysis, liver disease
steroid use, hyperthyroidism
Beta Hyperlipidemia, iron-deficiency Hypo-B-lipoproteinemia, malnutrition
 anemia
Polyclonal and Monoclonal Agammaglobulinemia,
Gamma
Gammopathies hypogammaglobulinemia

Monoclonal gammopathy

A dense narrow band that is composed of a single class of immunoglobulins secreted by an

abnormally expanded clone of plasma cells is known as M-protein (paraprotein, monoclonal
protein or M-component).[2] An M-protein usually presents as a single narrow peak, resembling a
"church spire," in the gamma, beta, or alpha-2 region of the densitometer tracing, or as a dense,
discrete band on the agarose gel (see image below).

Monoclonal pattern serum protein electrophoresis (SPEP).

The monoclonal antibody must be present at a concentration of at least 0.5 g/dL in order to be
accurately identified using SPEP. This corresponds to approximately 109 antibody-producing
cells.[3] Plasma cell disorders are typically associated with the presence of an M-protein (see
Table 2 below). In addition, an M component may be detected in other lymphoid malignancies
like chronic lymphocytic leukemia, any B- or T- cell lymphomas, breast cancer, colon cancer,
cirrhosis, sarcoidosis, and other autoimmune disorders.

Conditions associated with a monoclonal increase in the gamma region are as follows:

 Multiple myeloma

 Smoldering myeloma

 Monoclonal gammopathy of undetermined significance (MGUS)

  Waldenstrom macroglobulinemia

 Polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin

changes (POEMS syndrome)

 Solitary plasmacytoma

 Castleman disease

 AL amyloidosis

 Heavy chain deposition disease

 Light chain deposition disease

Polyclonal gammopathy

Infectious, Inflammatory or various reactive processes may be associated with a broad-based

peak or band in the gamma region (Figure 4). This pattern suggests a polyclonal increase in
immunoglobulins. Liver disease, autoimmune disease, chronic viral or bacterial infections and
various malignancies may cause a polyclonal rise in the gamma fraction (see Table 2 below).

Polyclonal pattern serum protein electrophoresis (SPEP).

Table 2. Diseases Associated with a Polyclonal Gammopathy (Open Table in a new window)

Condition Examples
Liver disease Cirrhosis, autoimmune or viral hepatitis
Rheumatoid arthritis, systemic lupus erythematosus,
Connective tissue diseases
scleroderma, Sjogren syndrome
Infection Bacterial: osteomyelitis, endocarditis, osteomyelitis
 Viral: HIV/AIDS, hepatitis C, Epstein-Barr virus

Hematologic Non-Hodgkin lymphoma, chronic lymphocytic leukemia,

disorders/malignancies thalassemia, sickle cell anemia
Nonhematologic malignancies Lung, ovarian, gastric malignancies, hepatocellular carcinoma

Considerations

The presence of an M-protein may be missed if the level is too small to be detected using SPEP.
In cases where a high suspicion for a clonal plasma cell disorder is high, more sensitive tests
such as a serum immunofixation or free light chain assay should be performed. In addition, when
either alpha-1 antitrypsin deficiency or immunoglobulin deficiency, specific quantitation is
indicated as SPEP is insensitive in these cases.

Collection and Panels

Specimen: Serum

Container: Red-top tube, serum separator tube (SST; see the image below)

Photograph of a red-top and serum-

separator (SST) tubes. (Original photo by Rapeepat Lekkham, MD.)

Collection method: Routine venipuncture

Blood must first be collected in a red top or serum separator tube. Serum, which is the fluid
portion of blood after it has clotted, is placed on a specific medium and exposed to an electric
current to separate its protein components. Two methods are usually used namely, agarose gel
electrophoresis, or less commonly, capillary zone electrophoresis.[2, 4, 5, 6] In both methods,
proteins are classified by their final position into five general regions:

Albumin

Albumin represents the highest peak in serum protein electrophoresis, usually seen as a single,
tall band. Occasionally, however two equally staining bands, referred to as bisalbuminemia or a
widely staining band may be seen, which both represent normal variations.[7] Common conditions
associated with decreased albumin include malnutrition, cirrhosis and nephrotic syndrome.
Dehydration, on the other hand, causes a high albumin.

An area between albumin and the alpha-1 band is called the albumin-alpha-1 interzone. Even
staining in this zone is caused by alpha-1 lipoprotein (High-density lipoprotein or HDL). An
increase is usually seen in alcoholic liver disease, pregnancy and during puberty. A sharp band
may also be seen in those with hepatocellular carcinoma as a result of elevated alpha-fetoprotein
(AFP).[1]

Alpha zone

The alpha-1 fraction includes alpha-1 antitrypsin, transcortin, and thyroid-binding globulin.[7]
Alpha-2 fraction is comprised of ceruloplasmin, alpha-2 macroglobulin, and haptoglobin. Both
alpha-1 and 2 represent the acute phase reactants; hence, malignancy, infection or any
inflammatory condition can cause their elevation. A relative increase in alpha-2 fraction may be
seen in nephrotic syndrome due to the relative larger size of the proteins and the inability to pass
through the glomeruli. A decrease in alpha-1 component may be seen in alpha-1 antitrypsin
deficiency and a decrease in alpha-2 component may be seen in hemolytic anemia due to
decreased haptoglobin levels.

Beta zone

The beta zone consist of beta-1 and beta-2 but is often represented a graphically as a single band.
Beta-1 consists mostly of transferrin, and is increased in conditions such as iron-deficiency
anemia, pregnancy and estrogen therapy. B-lipoprotein and C3 complement are included in the
B-2 component.[3] IgA, IgM, and sometimes IgG can occasionally be identified in the beta
fraction as well.

Gamma zone

Immunoglobulins mainly comprise this area including IgG, IgA, IgM, IgD and IgE.
Agammaglobulinemia and hypogammaglobulinemia syndromes such as IgA deficiency are
associated with a decrease in this area. Various inflammatory, autoimmune and hematologic, and
non-hematologic diseases are associated with an increased gamma peak. However, a
homogenous, spike-like increase in the gamma region is of special interest as it may represent an
abnormal expansion of immunoglobulin-producing plasma cells.
Background
Description

Serum protein electrophoresis (SPEP) is an easy, inexpensive method of separating proteins

based on their net charge, size, and shape. The 2 major types of protein present in the serum are
albumin and the globulin proteins. Albumin is the major protein component of serum and
represents the largest peak that lies closest to the positive electrode.[1] Globulins comprise a much
smaller fraction of the total serum protein but represent the primary focus of interpretation of
serum protein electrophoresis. Five globulin categories are represented: alpha-1, alpha-2, beta-1,
beta-2, and gamma, with the gamma fraction being closest to the negative electrode (see the
image below).

Normal serum protein electrophoresis diagram with legend of different zones.

Indications/Applications

Serum protein electrophoresis is generally considered in any patient with an elevated total
protein, especially those with elevated globulin level relative to albumin, or any signs and
symptoms suggestive of an underlying plasma cell disorder such as multiple myeloma,
Waldenstrom’s macroglobulinemia, or primary amyloidosis.[3] These include any of the
following:

 Unexplained anemia, back pain, bone pain, fatigue

 Unexplained pathologic fracture or lytic lesions

 Unexplained peripheral neuropathy

 Hypercalcemia secondary to possible malignancy

 Hypergammaglobulinemia

 Rouleaux formation noted on peripheral blood smear

 Renal insufficiency with bland urine sediment

 Unexplained proteinuria

 Bence Jones proteinuria

 Recurrent infections