Serum protein analysis and ag ratio
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sERUM PROTIENS AND AG RATIO, TOTAL PROTIENS, ALBUMIN, GLOBULINS, TOTAL ALBUMIN, SERUM GEL ELECTROPHORESIS
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Serum Protein and Albumin-Globulin Ratio
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Serum Protein and Albumin-Globulin Ratio
For MBBS Biochemistry Practical. Explains various methods of protein estimation and estimation of AG ratio, conditions leading to alterations in AG ratio etc.
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The document discusses serum protein electrophoresis, which separates serum proteins based on their charge and size. It notes that plasma is 92% water and contains 7.6 grams of protein per 100ml, including albumins, globulins, and fibrinogen. Albumin makes up 60% of total protein, globulins 35%, fibrinogen 4%, and other proteins 1%. The main types of globulins are also described. Serum protein electrophoresis is used to analyze the levels of specific proteins and identify abnormalities that can indicate underlying diseases or conditions.
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Plasma contains proteins that perform important functions like maintaining pH and colloid osmotic pressure. The most abundant protein is albumin, which makes up around 75% of plasma's colloid osmotic pressure. Other major classes of plasma proteins include globulins such as alpha-1 globulins containing thyroxine-binding globulin and alpha-1-antitrypsin; alpha-2 globulins containing ceruloplasmin and haptoglobin; beta-globulins containing transferrin; and gamma-globulins which are antibodies. These proteins transport substances like vitamins, minerals, lipids, and hormones or help in coagulation, immune defense, and acute phase responses.
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Plasma proteins have important functions including transport, osmotic regulation, catalytic functions, and protective functions. The major plasma proteins are albumin, globulins, and fibrinogen. Albumin is the most abundant plasma protein and serves important roles such as transporting metabolites, maintaining colloid osmotic pressure, and buffering. Electrophoresis is used to separate plasma proteins into fractions including albumin, alpha-1-globulin, alpha-2-globulin, beta-globulin, and gamma-globulin. Abnormal protein levels can provide clues for various clinical diseases.
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- The main plasma proteins are albumin, globulins, and fibrinogen. Albumin maintains osmotic pressure while globulins are involved in transport and immunity.
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Serum protein analysis and ag ratio
- 1. SERUM PROTEINS & A/G Ratio
- 2. • Plasma refers to the fluid portion of anticoagulant added blood after cells are removed by centrifugation • Serum is the fluid component of blood after it is allowed to clot
- 4. Albumin Most abundant plasma protein (~50%) Synthesized by hepatocytes Single polypeptide - 66 kDa High no. of charged AA results in ↑ solubility Plasma half life – 15 to 19 days Acts as a transporter and major component of colloid osmotic pressure (oncotic pressure)
- 5. Globulins Heterogeneous family of proteins Mol weight: 12-900 kDa Less water soluble than albumin Types Alpha (α) : α1 (α1-antitrypsin, α1-acid glycoprotein, HDL) α2 (Haptoglobin, α2-macroglobulin, Ceruloplasmin) Beta (β) : β1 (Transferrin,C4,LDL) β2 (C3, β2-microglobulin) Gamma (γ) : Immunoglobulins, C-reactive protein
- 6. Methods of Protein Estimation Copper based – Biuret assay (Biuret reagent) – Lowry assay (Folin-Ciocaltau reagent) – BCA assay (Bi-Cinchoninic Acid reagent) Dye based – Bradford assay
- 7. Principle Compounds with 2 or more peptide bonds, react with CuSO4 in alkaline conditions to produce a complex which is violet to blue colored.
- 10. Lowry assay
- 11. BCA assay
- 12. Bradford assay
- 13. Bromo-Cresol Green Method (Albumin analysis) • BCG reagent at pH 4.3 is negatively charged • pI of Albumin is 4.7 • Bromocresol green binds quantitatively with albumin to from an intense bluegreen • complex with an absorbance max at 670 nm.
- 17. A-G Ratio Direct: Albumin (A) & Globulins (G) in the serum sample are separated & determined separately Indirect: Serum total protein: Biuret reaction Serum albumin: Bromocresol Green (BCG) Serum globulin = Serum total protein - Serum albumin
- 18. Hypoproteinemia Hypoproteinemia is serum protein level < 6.7 g/dL ↓ (A/G) ↑ Loss • Renal disorders(e.g. nephrotic syndrome) ↓ Synthesis • Chronic liver disease(cirrhosis) ↑ (A/G) • Due to decrease in globulin • Agammaglobulinemia/ • hypogammaglobulinemia
- 19. Hyperproteinemia Hyperproteinemia is serum protein level > 8.3 g/dL Normal (A/G) • Relative hyperproteinemia • Dehydration ↓ (A/G) • Due to increase in globulin • Chronic infections • Malignant proliferation of plasma cells - e.g.myeloma
- 22. Liver damage Kidney damage
Editor's Notes
- Biuret Reagent Composition ;;Sodium hydroxide, Copper sulphate, Sodium potassium tartrate, Potassium iodide