Protein analysis

Protein in the serum is made up of albumin (∼ 60%) and globulin. Globulin is made up of alpha-1, alpha-2, beta, and gamma globulins. Although a total serum protein measures the total concentration of protein in the serum, it explains very little about the pathological etiology. Serum electrophoresis is an important diagnostic tool for the analysis of individual protein fractions which yields important information about underlying pathologies.

• Protein in the serum is made up of albumin (∼ 60%) and globulin. Globulin is made up of alpha-1, alpha-2, beta, and gamma globulins.
• A total serum protein (reference range 6–7.8 g/dL) test estimates the total of all protein fractions together, e.g. Biuret test
  • Albumin: 3.5–5.7 g/dL (35–57 g/L)
  • Globulin
    • alpha-1: 0.1–0.4 g/dL (1-4 g/L)
    • alpha-2: 0.3–0.9 g/dL (3–9 g/L)
    • beta: 0.7–1.5 g/dL (7–15 g/L)
    • gamma: 0.5–1.4 g/dL (5–14 g/L)
• If the result is not within the reference range, one of the following tests is usually performed for further analysis:
  • Quantitative analysis of single proteins: immunohistochemical tests (see immunohistology)
  • (Semi‑)qualitative analysis of different protein fractions: serum electrophoresis (see below)

References:^[1][2][3]

• Definition: laboratory method to separate proteins and their fragments according to size and electrical charge.
• After staining and visualization of the results, characteristic patterns associated with certain pathologies become visible.

Indications

• Suspected multiple myeloma
• Light-chain amyloidosis
• Unexplained peripheral neuropathy
• Suspected malignancy
• Renal insufficiency

Procedure

1. Load serum sample into agarose gel and apply an electrical current
2. Proteins migrate through the gel
3. Any proteins with similar properties (size, charge) will accumulate and form bands, regardless of their individual functions
4. These bands are stained, enabling them to be analyzed and visualized under UV light and charted in an absorption curve.
5. The relative quantity of certain proteins can be determined by calculating the area under the curve.
6. Absolute concentrations of serum proteins can be calculated by multiplying the percentage of a respective fraction by the total serum protein concentration.

Analysis

References:^{[2][4][5][6][7][8][9][10][11][12]}