an anticoagulant Contains fibrinogen + albumin and globulins SERUM The fluid portion of the blood obtained after removal of the fibrin clot and blood cells SERUM=PLASMA-FIBRINOGEN
CONTENTS OF PLASMA PROTEIN 0.9% inorganic ions, 0.8% small organic molecules and 7% protein Total protein content of N.plasma 6 -8 gm/dl Albumin 3.5 -5 gm/dl Globulins - 2.5 -3.5gm/dl Fibrinogen 200-400mg/dl Alb : glb ratio -1.2:1 to 1.5:1
CHARACTERICTIC OF PLASMA PROTEIN Most of them are synthesized in the liver Albumin and fibrinogen; 50 - 80% Globulin Rest of the globulin Lymphoid tissue Almost all are glycoproteins Each one has a characteristic half-life in the circulation
Plasma protein Specific plasma protein Albumin Transport protein Acute phase protein Immunoglobulins Complement protein Cytokines Coagulation factors Plasma enzymes
ALBUMIN Most abundant plasma protein Synthesized by hepatocytes used in LFT A low molecular weight protein (69,000D) Significant component of most ECF Single polypeptide chain 585 amino acids with 17 intra chain S-S bonds Highly soluble in water Half life 15-19 days
Role in Transport: Transports metals, ions, fatty acids, amino acids, bilirubin, enzymes, drugs etc.. Maintainance of colloidal osmotic pressure Buffering action
FUNCTIONS OF ALBUMIN Maintenance of colloidal oncotic pressure Proteins exert effective osmotic pressure 25mm Hg; 80% contributed by Albumin STARLINGS HYPOTHESIS : At the capillary end; BP / Hydrostatic pressure expels water out Effective osmotic pressure takes water into the vascular compartment
CAUSE FOR EDEMA ?.... Arterial end of capillary BP - 35mm Hg EOP - 25 mm Hg Thus water expelled by a P of 10 mm Hg Venous end - BP - 15mm Hg EOP - 25 mm Hg Thus water is imbibed with P of 10 mm Hg Therefore blood volume remains the same If S.protein reduces;EOP decreases; leads to accumulation of water
CAUSES OF HYPOALBUMINAEMIA DECREASE IN ALBUMIN SYNTHESIS Cirrhosis of liver disease (chronic diseases) Malnutrition Haemodilution a. Over hydration b. Late stage of pregnancy C. Artefactual-Blood drawn from driparm
INCREASED ALBUMIN SYNTHESIS Nephrotic syndrome Extensive burns Malabsorption due to gastro- intestinal disease Protein-losing enteropathy a. Ulceration of the bowel b. Lymphatic obstruction
ALPHA ALBUMIN
1 is comprised : 1-antitrypsin
1- acid glycoprotein
Alpha fetoprotein
1- lipoprotein HDL 2 consist of:
Ceruloplasmin 2-macroglobulin Haptoglobin- binds to hemoglobin
Ceruloplasmin 2 globulin;contains 95% of total copper found in serum Single polypeptide chain Transport protein for Cu; Acute phase protein Synthesized in liver Copper is absorbed and transported to liver but absence of hepatocellular Ptype ATPase prevents incorporation of Cu into Cp
Wilsons disease Hepatolenticular degenaration Autosomal recessive Mutation in gene coding for copper binding ATPase(excretion of Cu from cells) Decrease in Cp levels Marked increase in tissue copper liver,brain,kidney Increased urinary Cu and tissue Cu
Menkes disease X-linked inherited disorder Dietary Cu absorbed but cannot be transported to blood due to genetic absence of an intracellular Cu binding ATPase Cu accumulates affects vascular and connective tissues
Beta and gamma- globulins Beta-globulin: Haemopexin Transferrin C-reactive protein 2-macroglobulin LDL lipoprotein
Gamma-globulin: Immunoglobulins Ig G, Ig M, Ig A, Ig D,Ig E Transferrin Principal plasma transport protein for iron Plasma levels regulated by levels of iron One molecule binds 2 molecules of iron Fe-Tf complex is taken up by all tissues by specific surface receptors
Lipoproteins Function to transport cholesterol, TAG & phopholipid in blood Sub classified into VLDL, IDL, LDL, HDL
Acute phase response and proteins Nonspecific response to tissue injury or infection Assessment of the presence and degree of inflammation Positive APPs: Increase in inflammation Negative APPs: Decrease in inflammation Presence of inflammatory disease Monitoring therapeutic effectiveness Follow-up of patient with malignancy
Acute phase reactants Positive acute phase proteins C-reactive protein 1-antitrypsin 1-antichymotrypsin 1-acid glycoprotein Ceruloplasmin Haptoglobin Complement component C3 and C4 Negative acute phase proteins Albumin Transferrin Pre-albumin
Estimation of serum proteins Total protein estimation: Biuret Method Colorimetric method Most widely used Principle: Alkaline Cu 2+ reacts with compounds containing 2 or more peptide bonds to give a violet-colored complex, which is then measured at 540 nm Analysis of proteins Specific quantitative assays- immunochemical methods nephelometry turbidometry electroimmunoassay RIA, fluorescence immunoassay chemiluminescence Structural and quantitative information - Mass specrometry
Estimation of s.proteins.. Estimation of serum albumin Salt precipitation: Globulins are precipitated in high salt concentration; albumin in supernatant is quantitated by biuret reaction Dye binding: Albumin binds to dye; causes a shift in the absorption maximum BCG (bromocresol green) BCP (bromocresol purple)
Electrophoresis Technique invented by Tiselius in 1937,Nobel prize in 1948 Proteins separated based on electric charge The process of moving charged molecules in solution by applying an electrical field across the mixture Molecules move with a speed dependent on their charge, shape, and size
PLASMA PROTEIN Total serum protein Amount in grams per deciliter (g/dL) Amount in SI units grams per liter (g/L) Albumin 58%74% 3.55.5 3555 Alpha-1 globulin 2.0% 3.5% 0.20.4 24 Alpha-2 globulin 5.4% 10.6% 0.50.9 59 Beta globulin 7%14% 0.61.1 611 Gamma globulin 8%18% 0.71.7 717
Serum Total protein - 6.4 to 8.3 g/dL Albumin - 3.5 to 5.0 g/dL Serum globulin - 2.5 to 3.5 g/dL
Paraprotein Is an abnormal plasma protein appearing in large quantity; ELOMA Malignant proliferation of plasma cells; S. Ig G /Ig A Multiple myeloma Light chains(low mol wt.) present in urine are considered as Bence Jones proteins Starts precipitating on heating urine at 45 - 60