Plasmatic proteins.

Protein electrophoresis.
Albumin

• is the major protein of human plasma

• makes up approximately 60% of the total plasma protein
• about 40% of albumin is present in the plasma and the other 60% is
present in the extracellular space
• albumin is synthesized in the liver
Functions of Albumin
• colloidal osmotic pressure (due to its low molecular wight and large
concentration)→ is responsible for maintaining the blood fluids from
leaking out from intravascular to extravascular space

• transports molecules that are hydrophobic:

-metals (calcium, copper, heavy metals)
-FFA
-bilirubin (albumin-bilirubin complex)
-hormones
-drugs (sulfonamides, penicillin G, dicoumarol)
Clinical significance of Albumin
Reference range: 4,5 – 5,5 g/dl
Causes of hypoalbuminemia Causes of hyperalbuminemia

Physiological: Pathological: -acute dehydration

pregnancy 1. Impared synthesis:

-malnutrition
-malabsorption
-liver disease

2. Abnormal distribution:
-burns

3. Excessive losses:
-nephrotic syndrome
-protein-lose entheropathy

4. Increased catabolism:
-malignant disease

Hypoalbuminemia → fall of plasma osmotic pressure → retention of fluid in the tissue spaces = Hyperalbuminemia → no clinical significance
EDEMA
Globulins
• high molecular weight
• their electrophoretic migration rates is lower than for albumin
• α and β globulins are synthesized in the liver
• γ globulins are synthesized in:
-plasma cells
and
-B-cells of lymphoid tissues (reticulo-endothelial system)
Functional classification:
• Plasmatic inhibitors of proteases:
-α1-antitrypsin
-α1-chymotrypsin
-α2-macroglobulin
-α2-antiplasmin
• Acute phase proteins: CRP, PCT, haptoglobin, ceruloplasmin, α1-acid
glycoprotein
• Proteins with transporting role: apoB, apoE; ferritin, transferrin;
ceruloplasmin; haptoglobin, myoglobin
• Coagulation factors:
-the intrinsic pathway factors: I, II, V, VIII, IX, X, XI, XII
-the extrinsic pathway factors: VII
• Oncofetal proteins:
-α1-fetoprotein
-β2-microglobulin
Alfa 1 globulins
• α1-antitrypsin
• α1-acid glycoprotein
• α1-lipoprotein (HDL) – transport of lipids
• α1-thyroid-binding globulin – transport of thyroid hormones
• α1-transcortin – transport of cortisol, corticosterone and progesterone
• α1-fetoprotein
Alfa 2 globulins
• α2-macroglobulin
• Haptoglobin
• Ceruloplasmin – transport of copper
• Retinol-binding protein – transport of vitamin A
• Vitamin D-binding protein – transport of Cholecalciferol
Alfa globulins
α1 - globulins α2 - globulins
α1-ANTITRYPSIN α2-MACROGLOBULIN
= α1-antitprotease -is the major component of the the α2 fraction of human plasma
-is the major component of the α1 fraction of human plasma -it is synthesized by hepatocytes and macropahges
-it is synthesized by hepatocytes and macropahges -is the principal proteolytic enzymes inhibitor (it inhibits coagulation enzymes,
-is the principal proteolytic enzymes inhibitor (it inhibits proteases, trypsin plasmin, elastase)
and elastase) -↑ levels in protein losing state (nephrotic syndrome)
-↑ levels in acute inflamation -the genetic defect is incompatible with life !!
α1-ACID GLYCOPROTEIN CERULOPLASMIN
=oroscomucoid -carrier for Copper (90%)
-it is synthesized by the liver parenchymal cells -↑ levels in acute inflamation
-it contains a high percentage of carbohydrate that are transported to the site -↓ levels in Wilson disease (abnormal metabolism of copper)
of tissue injury
-↑ levels in acute inflamation
α1-FETOPROTEIN HAPTOGLOBIN
=AFP -it binds with Hb to form a complex which is too large to be filtred by the
-it is synthesized during pregnancy in the tissues and plasma of the fetus kidney → prevents the loss of iron in the urine
-Adults: <1µg/dl -↑ levels in acute inflamation
-↑ levels in hepatocellular carcinoma or teratoblastomas -↓ levels in hemolytic anemias
Beta globulins
• CRP
• Fibrinogen
• Hemopexin – transport of heme
• Transferin – transport of iron
• Transcobalamin – transport of vitamin B12
• β-lipoprotein (LDL) – transport of lipids
• β2-macroglobulin: ↑ levels in myeloma patients and those with renal
failure
Gama globulins
• IgA – (made in lamina propria of intestinal and laminal tract):
mucosa-protecting antibodies
• IgG – (transferred to baby from mothers blood across the placenta):
protects the extravascular tissues spaces - late antibodies
• IgM – (foetus can synthesize IgM but levels are low at birth so a ↑
level at birth indicate an intrauterine infection) - protects the blood
stream against foreign antigens - early antibodies
• IgD – ??
• IgE – (produced by plasma cells in respiratory tract) - ↑ allergic
reactions
Total serum proteins: 6-8 g/dl
Albumin: 4,5 – 5,5 g/dl
Globulins: 1,5 – 2,5 g/dl
Albumin: 52-59%
α1 – globulins: 3-5%
α2 – globulins: 7-9%
β – globulins: 11-14%
γ – globulins: 16-20%
Clinical significance
Modifications in the protein levels:

Hyperproteinemia
-monoclonal gammapathies: multiple myeloma, Waldenstrom
macroglobulinemia

Hypoproteinemia
-affected synthesis of proteins - liver diseases: ?
-loss of proteins through the kidneys - kidney diseases: ?
Clinical significance
Modifications of the protein fractions:

1. Acute inflammation
2. Chronic inflammation
3. Multiple myeloma
4. Hepatic cirrhosis
5. Nephrotic syndrome
6. Hypogammaglobulinemia
Acute inflammation
Chronic inflammation
Multiple myeloma
Hepatic cirrhosis
Nephrotic syndrome
Hypogammaglobulinemia
Principle of plasma total proteins determination:

Reagents Sample Standard Blank

Serum 1 ml
Protein standard 1 ml
Distilled water 1 ml
Gornall reagent 4 ml 4 ml 4 ml
Mix the tubes and incubate 15 min. at room temperature.

Measure the absorbances ( As, Ast) at 540 nm.

𝐀𝐬 𝐀𝐬
Calculation: mg total proteins/ml = × 𝐂𝐬𝐭 = × 𝟏𝟎
𝐀𝐬𝐭 𝐀𝐬𝐭