DR.LAXMIKANTA SAY

PLASMA PROTEIN

1910- Albrecht Kosel a German scientist got nobel prize for the work “ in recognition of the contribution to our knowledge of cell chemistry made through work on proteins , including the nucleic substances.

HISTORY

American wounded soldier receiving blood plasma in August, 1943 ."Dried plasmas" were developed and first used in World War II. Prior to the United States' involvement in the war, liquid plasma and whole blood were used. The "Blood for Britain" program during the early 1940s was quite successful (and popular in the United States) based on

Dr. Charles Drew's contribution. The resulting dried plasma package came in two tin cans containing 400 cc bottles. One bottle contained distilled water to reconstitute the dried plasma. In about three minutes, the plasma ready to use, stay fresh for around four hours

HISTORY

PLASMA

SOLIDS(9%)

INORGANIC(1%)

ORGANIC(8%)

WATER(91%)

Extracellular-Na+, Ca++,Cl-,HCO3-

Intracellular-K+, Mg++, Cu++, PO4---,Protein-

Others-Fe++, Fe+++,

INORGANIC MOLECULES

ORGANIC MOLECULESPlasma

protein- 7% -Albumin-55% -Globulin-38% -Fibrinogen-7% -Prothrombin

Non-protein Nitrogen (NPN)- 1%

-Urea :20-40 mg/dL-Uric acid : 2-4 mg/dL-Creatine :1-2 mg/dL-Creatinine :0.6-

1.2mg/dL-Xanthine :traces-Hypoxanthine :traces

Neutral fats (Triglycerides) : 30-150 mg/dL

Phospholipid :Lecithin ,spingomyelin, cephalin

Glucose (fasting) : 70-90 mg/dL

Cholesterol :120-200 mg/dL

OTHERS

Coagulation of bloodMaintain colloidal osmotic pressureMaintaining viscosity of bloodMaintaining systemic arterial pressureStability to bloodMaintain acid-base balanceImmunity Transport functionReservoir function

FUNCTIONS

55% blood volume , 5% B.W.Types : Prealbumin & AlbuminPrealbumin : MW-60,000, Normal plasma level-0.03 gm/dL , -binds Thyroxine (T 4 and T3)Albumin : MW69,000, Normal plasma level-3-5 gm/dL (avg. 4.8 gm/dL)o-Controls colloidal osmotic pressure-Binding and Carrier protein : anions, cations ,dyes, drugs, hormones, fatty acids, metals, aminoacids , enzymes & bilirubin

ALBUMIN

GLOBULIN •Can be separated into different fractions on the basis of their electrophoretic mobility and sedimentation coefficient:

α1-Globulin-α1-Fetoprotein

-α1-Antitrypsin

α2-Globulin-α2-Fetoprotein

-Haptoglobin

β-Globulin-Transferrin -

Ceruloplasmin γ-Globulin-Antibodies

(immunoglobulins)

Types :-α –globulin – 0.78-

0.81 gm/dL-β- 0.79-0.84

gm/dL-γ-globulin -0.66-

0.70gm/dL

Precipitation by salt

Sedimentation in Ultracentrifuge

Electrophoresis

Isoelectric point

METHOD OF STUDY

Physical Techniques1.Ultracentrifugation (analytical or Sedimentation velocity ultracentrifuge) at 60,000 per.min. (Refractive index the boundary between the solvent and the protein is visualized by an optical system –called Sehlieren System).AdvantageMost useful for the determination of the mol. wt of proteinsDisadvantageHigh cost of each analysis and poor resolving capacity (when applied to whole serum or plasma)

ULTRA CENTRIFUGATION

Protein in aqueous solution are charged groups (e.g. carboxylic (Asp.Glu), amino groups (Lys,Arg), they can be separated under an electric field using various stabilizing media. N.B. Amino groups undergo ionic dissociation at alkaline pH and carboxylic undergo dissociation at acid pH. Most proteins are –ve at pH 8.6. The pH at which +ve charges equal to –ve charges is characteristic for a protein and is called isoelectric point ).•Boundary electrophoresis: Separation in free liquid media•Zone electrophoresis -Separation in stabilizing media(e.g. Paper, Cellulose acetate, Starch, Polyacrylamide , Agarose)

ELECTROPHORESIS

Separates proteins on the basis of their charge.-Types:-Free boundary: separation under an electric field in a fluid media. Separates plasma proteins five bands: albumin(54-58%), α1 globulins(6-7%), α2 globulins( 8-9%), β -globulins (13-14%),γ-globulins (11-12%).-Zone electrophoresis: Separation under an electric field in a solid media e.g. paper, starch, cellulose, Acrylamide etc. Separates plasma proteins into: Albumin, α1 globulins, α2 globulins, βglobulins,γglobulins and fibrinogen.

ELECTROPHORESIS

1.Physiological :a. Newborn and infancyb. Pregnancy2.Excessive loss of protein:a. Through the kidney in nephrotic syndromeb. From the skin after burnsc. Through the skin in protein losing enteropathy.3.Decreased synthesis of proteinsa.Dietary protein deficiency in Kwashiokarb.Liver disease (Hepatitis, Cirrhosis).c. Malabsorption.

HYPOPROTEINEMIA

HYPOALBUMINAEMIA Decrease albumin

synthesis:-a. Liver disease (chronic

diseases).-b. Malnutrition.-c. Alcoholism•Increased albumin

loss:-a. Renal disease

(nephrotic syndrome).-Loss of albumin in urine

(proteinuria).-b. Extensive burns:-Loss of albumin through

skin -transdution.

Defective intake:-a. Malabsorption due to

gastro-intestinal disease .

-Protein-losing enteropathy

–Excessive loss of protein from the body into the gut.

- Conditions such as :a. Ulceration of the

bowel.b. Lymphatic

obstruction.c. Intestinal lymphaangiectasis.

Multiple myeloma

T.B.

Lymphatic leukemia

Cirrhosis of liver & Acute hepatitis

Nephritis

INCREASED γ-GLOBULIN

FIBRINOGEN DECREASED-Congenital-Carcinoma prostrate-Extensive cardiac &

pulmonary surgeries

-Intravascular coagulation

INCREASED-Pregnancy,

menstruation-Malaria-Tissue injury-acute & chronic

infections

ACUTE PHASE PROTEIN

Indicates active stage of inflammation Differential diagnosis of inflammatory

disease. Estimation of the endpoint of therapy. Monitoring therapeutic effectiveness. Post surgical follow-up in patients at

risk of postoperative infections. Follow-up of patient with malignancy.

•C-reactive protein•α1-antitrypsin.•α1-antichymotrypsin.•α1-acid glycoprotein.•Ceruloplasmin.•Haptoglobin.•Complement component C3 and C4.•Antithrombin III.

PROTEIN DISEASEα1-antitrypsin -Chronic obstructive pulmonary disease. -Neonatal hepatitis syndrome , -cytogenic cirrhosis.Ceruloplasmin- Hepatitis or cirrhosis Haptoglobin -- - In-vivo haemolysis. -Ineffective erythropoiesisα1-Antitrypsin -Obstructive pulmonary disease (Chronic or emphysema) liver disease.Anti-thrombin-Thrombosis -Pulmonary embolismImmunoglobulin -Severe recurrent Complement-Recurrent infection.C1 esterase inhibitor - Recurrent non- pruritic swelling of skin and mucus membrane (hereditary angioneuroticedema

Abnormal collection of fluid in interstitial spaces.Decrease in plasma levelDecreased COPIncreased filtration at arterial level & decreased absorption at venous end .When capillary permeability increased e.g.in anorexia, urticaria, inflammation etc,-protein escapes from capillary membrane interstitial spaces producing oedema

OEDEMA

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