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Questions and Answers
What primarily determines the speed of protein running in gel electrophoresis?
What primarily determines the speed of protein running in gel electrophoresis?
What is the role of a densitometer in serum protein electrophoresis?
What is the role of a densitometer in serum protein electrophoresis?
Which protein is most commonly found in the Albumin Zone during serum protein electrophoresis?
Which protein is most commonly found in the Albumin Zone during serum protein electrophoresis?
What condition may cause a decrease in albumin levels observable on electrophoresis?
What condition may cause a decrease in albumin levels observable on electrophoresis?
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What is the primary role of albumin in plasma?
What is the primary role of albumin in plasma?
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What does an increased level of Alpha-1 antitrypsin indicate?
What does an increased level of Alpha-1 antitrypsin indicate?
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Which of the following proteins is responsible for blood clotting?
Which of the following proteins is responsible for blood clotting?
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Which of the following proteins is primarily associated with the Alpha-2 zone?
Which of the following proteins is primarily associated with the Alpha-2 zone?
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What percentage of plasma is made up of water?
What percentage of plasma is made up of water?
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What indicates a low level of haptoglobin in serum protein electrophoresis?
What indicates a low level of haptoglobin in serum protein electrophoresis?
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Which type of globulin constitutes the largest portion of globulins and consists mostly of immunoglobulins?
Which type of globulin constitutes the largest portion of globulins and consists mostly of immunoglobulins?
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What happens to alpha-2 macroglobulin levels during inflammation?
What happens to alpha-2 macroglobulin levels during inflammation?
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Where are 90% of plasma proteins synthesized?
Where are 90% of plasma proteins synthesized?
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What is the primary function of serum protein electrophoresis (SPEP)?
What is the primary function of serum protein electrophoresis (SPEP)?
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What happens to serum proteins at a pH of 8.6 during electrophoresis?
What happens to serum proteins at a pH of 8.6 during electrophoresis?
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Which of the following is NOT classified as a plasma protein?
Which of the following is NOT classified as a plasma protein?
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What is the primary reason for the significant increase in AMG during nephrotic syndrome?
What is the primary reason for the significant increase in AMG during nephrotic syndrome?
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Which zone of protein electrophoresis contains transferrin and beta-lipoprotein?
Which zone of protein electrophoresis contains transferrin and beta-lipoprotein?
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Which protein is typically increased in the acute phase response?
Which protein is typically increased in the acute phase response?
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In which condition would you expect an increase in beta-1 protein due to free transferrin?
In which condition would you expect an increase in beta-1 protein due to free transferrin?
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What is the visible result when blood is drawn from patients treated with heparin?
What is the visible result when blood is drawn from patients treated with heparin?
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What is a characteristic appearance of a normal gamma region on protein electrophoresis?
What is a characteristic appearance of a normal gamma region on protein electrophoresis?
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What contributes to the decreased level of complement protein 3 (C3) in autoimmune disorders?
What contributes to the decreased level of complement protein 3 (C3) in autoimmune disorders?
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What defines the structure of immunoglobulins?
What defines the structure of immunoglobulins?
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What is a characteristic change in the acute inflammatory response?
What is a characteristic change in the acute inflammatory response?
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Which statement correctly describes chronic inflammation?
Which statement correctly describes chronic inflammation?
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In cases of liver damage such as cirrhosis, which of the following changes is typically observed?
In cases of liver damage such as cirrhosis, which of the following changes is typically observed?
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What is a common feature of nephrotic syndrome?
What is a common feature of nephrotic syndrome?
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Monoclonal gammopathy is most commonly associated with which condition?
Monoclonal gammopathy is most commonly associated with which condition?
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Which of the following proteins are classified as negative acute-phase proteins?
Which of the following proteins are classified as negative acute-phase proteins?
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Which laboratory finding is typically present in hypergammaglobulinemia?
Which laboratory finding is typically present in hypergammaglobulinemia?
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In acute inflammation associated with infection or injury, what happens to albumin levels?
In acute inflammation associated with infection or injury, what happens to albumin levels?
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What is a significant deleterious effect of increased protein levels in the body?
What is a significant deleterious effect of increased protein levels in the body?
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What does a spike in serum protein electrophoresis indicate?
What does a spike in serum protein electrophoresis indicate?
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Which type of monoclonal protein is associated with Bence Jones protein?
Which type of monoclonal protein is associated with Bence Jones protein?
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What type of immunoglobulin was identified in the patient's case of multiple myeloma?
What type of immunoglobulin was identified in the patient's case of multiple myeloma?
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What does immunofixation electrophoresis (IFE) help further identify?
What does immunofixation electrophoresis (IFE) help further identify?
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How is monoclonal gammopathy commonly diagnosed through serum protein analysis?
How is monoclonal gammopathy commonly diagnosed through serum protein analysis?
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In the diagnosis of multiple myeloma, what important finding is typically observed in the bone marrow?
In the diagnosis of multiple myeloma, what important finding is typically observed in the bone marrow?
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What happens during the precipitation reaction in immunofixation electrophoresis?
What happens during the precipitation reaction in immunofixation electrophoresis?
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Study Notes
Clinical Chemistry Case Study
- This study examines serum protein electrophoresis (SPEP) and respiratory system history and examination, focusing on a₁-antitrypsin deficiency.
Introduction to Plasma
- Plasma comprises 46-63% of blood.
- It appears straw-colored or clear, similar to interstitial fluid.
- Key differences include respiratory gas levels and dissolved protein concentration/types.
- Plasma composition is about 92% water.
- Proteins in plasma (per 100 ml):
- Albumins (60%)
- Globulins (35%)
- Fibrinogen (4%)
- Other proteins (1%)
Serum Proteins
-
Albumin:
- Most abundant plasma protein.
- Water-soluble, produced by the liver.
- Maintains plasma osmotic pressure.
-
Globulins:
- Four types: alpha 1, alpha 2, beta, and gamma.
- Diverse functions, including transport of substances and the formation of other substances.
- Gamma globulins are a major portion, with many being immunoglobulins.
-
Fibrinogen:
- A clotting protein produced by the liver.
- Converts to fibrin during clotting.
- Absent in standard serum protein electrophoresis.
-
Other Plasma Proteins:
- Remaining 1% of plasma consists of various peptide hormones (e.g., insulin, prolactin) and glycoproteins (e.g., TSH, FSH, LH).
Plasma Protein Origins
- Liver synthesizes about 90% of plasma proteins.
- Lymphocytes (lymphatic system) produce plasma cells which synthesize antibodies.
- Endocrine organs produce peptide hormones.
Serum Protein Electrophoresis (SPEP)
- A laboratory technique to identify protein types.
- Venous blood samples are processed into serum,
- Separates proteins by size and charge (typically into 4-6 fractions, sometimes 2 additional beta fractions).
- A useful diagnostic tool for various diseases.
Serum Protein Electrophoresis (SPEP) Principle (Agarose Gel)
- Serum proteins at pH 8.6 are negatively charged.
- A buffer maintains constant pH.
- Protein migration toward the anode is determined by net charge and molecular weight.
- Separated proteins are fixed and stained for visualization.
SPEP Diagram
- Shows six bands: albumin at the beginning, followed by a₁-, a₂-, β₁-, β₂-, and γ-globulins running from front to back, in the order given.
Protein Electrophoresis Use in Diagnostics
- Serum protein bands are quantified to aid disease diagnosis.
- Densitometry uses image processing software to analyze separated proteins' concentrations.
- Reference ranges are used for comparisons.
- Specific proteins (e.g., alpha 1-antitrypsin) serve as markers for individual conditions.
Major Contents of Electrophoresis Bands
- Alpha 1: Alpha 1-antitrypsin.
- Alpha 2: Alpha-macroglobulin and haptoglobin.
- Beta: Beta-lipoprotein (LDL), transferrin.
- Gamma: Immunoglobulins (IgG, IgM, IgA, IgD, IgE)
Albumin Zone
- Albumin is the major fraction in normal SPEP.
- A 30% decrease is typically needed to be visually notable.
- Usually seen as a single band near the start.
- Low levels suggest liver disease, malnutrition, malabsorption, or protein loss conditions.
Albumin - alpha-1 interzone
- High alpha fetoprotein (AFP) levels can produce a sharp band between albumin and alpha-1, suggesting hepatocellular carcinoma.
Alpha-1 Zone
- Alpha-1 antitrypsin (A1AT) is primary in this band.
- A decrease in this zone is seen in alpha-1 antitrypsin deficiency.
- Other proteins migrate in this zone (e.g., alpha-lipoprotein, orosomucoid).
Decreased Alpha 1 Antitrypsin
- Associated with conditions such as chronic liver disease (hepatic cirrhosis).
- Protein loss (e.g., nephrotic syndrome) also contribute to decreases.
Increased Alpha 1 Anti-Trypsin
- Elevated levels likely suggest acute inflammation.
Alpha-2 Zone
- Principally consists of alpha-2 macroglobulin (AMG) and haptoglobin.
- Haptoglobin levels are low in hemolytic anemia (as haemoglobin is lost).
- Haptoglobin levels rise in acute inflammation.
Increased Alpha-2 Macroglobulin
- Elevated levels often indicate nephrotic syndrome (protein loss).
Beta Zone
- Divided into Beta 1 and Beta 2.
- Beta 1 subzone includes transferrin and beta-lipoprotein.
- Increased beta-1 protein is linked with iron deficiency anemia, pregnancy, estrogen therapy, and hypercholesterolemia.
Beta-2 Zone
- Predominantly consists of Complement protein 3 (C3) and fibrinogen.
- Increased C3 levels are related to the acute phase response.
- Decreased levels in autoimmune disorders.
- Fibrinogen is absent in normal serum, occasionally shown as a band in undiluted serum.
Gamma Zone
- Primarily composed of immunoglobulins (IgG, IgM, IgA, IgD, and IgE).
- A smooth 'blush' or smear, free of asymmetry or sharp peaks, characterizes the normal zone.
- Elevated or decreased levels, or abnormal peaks, can indicate various health conditions.
- Immunoglobulin subtypes may be identified by immunofixation electrophoresis.
Immunofixation Electrophoresis (IFE)
- IFE further identifies abnormal protein bands, revealing specific immunoglobulin types.
- Antigens (immunoglobulins) precipitate when interacting with antibodies for specific visualization.
- A useful technique to verify monoclonal gammopathy presence.
Monoclonal Gammopathy
- Characterized by high amounts of a single type of abnormal immunoglobulin.
- Often linked with immunoproliferative disorders (e.g., multiple myeloma).
- The excess immunoglobulin has harmful impacts, such as elevated blood viscosity and kidney damage.
- Immunofixation electrophoresis (IFE) is often employed for diagnosis.
Summary of SPEP in Different Disease States
- Different disease conditions will show distinct patterns in SPEP tests.
Acute Inflammatory Response
- Characterized by elevated a1 and a2 globulins.
Chronic Inflammatory Response
- Associated with normal or decreased albumin and elevated a1 and a2 globulins, while showing increased γ-globulins.
Liver Damage - Cirrhosis
- Shows decreased albumin and a1, a2, and β globulins, with a noticeable increase in γ-globulins.
Nephrotic Syndrome
- Associated with decreased albumin and increased a2-macroglobulin.
Hypogammaglobulinemia
- Associated with decreased γ-globulins.
Polyclonal Gammopathy
- Characterized by diffusely elevated γ-globulin, commonly linked with liver disease, chronic inflammation, or malignancy.
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Test your knowledge on serum protein electrophoresis with this engaging quiz. Explore questions about the roles and levels of different proteins, their functions, and the techniques used in this analytical method. Perfect for students and professionals in biochemistry and laboratory sciences.
