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Protein analysis

Last updated: December 6, 2019

Summarytoggle arrow icon

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:

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

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

Name of protein fraction

Albumin

Alpha-1 globulins Alpha-2 globulins Beta globulins Gamma globulins
Fraction contains
Increased in
Decreased in
  • Alpha-1 antitrypsin deficiency
  • Liver disease

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

  1. Devaraj S. Albumin . In: Wheeler TM, Albumin . New York, NY: WebMD. http://emedicine.medscape.com/article/2054430. Updated: January 13, 2015. Accessed: May 9, 2017.
  2. Alvaran Tuazon S. Serum Protein Electrophoresis. In: Staros EB, Serum Protein Electrophoresis. New York, NY: WebMD. http://emedicine.medscape.com/article/2087113. Updated: September 5, 2014. Accessed: May 9, 2017.
  3. Chauhan DBS. Principles of Biochemistry and Biophysics. Firewall Media ; 2008
  4. Gel Electrophoresis Overview. http://scienceprimer.com/gel-electrophoresis-overview. Updated: January 1, 2017. Accessed: May 9, 2017.
  5. What is Gel Electrophoresis?. http://www.yourgenome.org/facts/what-is-gel-electrophoresis. Updated: January 25, 2016. Accessed: May 9, 2017.
  6. Lee MT, Caturegli P, Humphrey RL, Thompson RE, Detrick B. γ/IgG ratio: Role in distinguishing monoclonal spikes from fibrinogen. J Clin Lab Anal. 2011; 25 : p.332-336.
  7. Effect of hemolysis on serum protein electrophoresis.
  8. Alberghina D, Giannetto C, Vazzana I, Ferrantelli V, Piccione G. Reference intervals for total protein concentration, serum protein fractions, and albumin/globulin ratios in clinically healthy dairy cows. J Vet Diagn Invest. 2011; 23 (1).
  9. Colman RW. Hemostasis and Thrombosis: Basic Principles and Clinical Practice. Lippincott Williams & Wilkins ; 2006
  10. Rajkumar SV. Laboratory Methods for Analyzing Monoclonal Proteins. In: Post TW, ed. UpToDate. Waltham, MA: UpToDate. https://www.uptodate.com/contents/laboratory-methods-for-analyzing-monoclonal-proteins.Last updated: October 14, 2015. Accessed: May 9, 2017.
  11. O'Connell TX, Horita TJ, Kasravi B. Understanding and interpreting serum protein electrophoresis. Am Fam Physician. 2005; 71 (1): p.105-112.
  12. Shah R. General Practice. PasTest Ltd ; 2005
  13. Herold G. Internal Medicine. Herold G ; 2014
  14. Protein. http://medical-dictionary.thefreedictionary.com/protein. . Accessed: May 9, 2017.