Blood Components and Plasma derivatives

M. Zaharna Blood Bank Lab. 20091

Blood Components

and Plasma derivatives


• The anticoagulants and preservatives that are added to blood nowadays enable storage for long periods of time– Before acid-citrate-dextrose (ACD) was used- 21 days

• Aseptic separation of blood into cellular & plasma components by the introduction of plastic collection systems– Before glass bottles were used


Blood collection

• Blood is collected in plastic bag systems with anticoagulant & preservative

• Whole blood can be stored at 4oC for up to 5 weeks

• Whole blood contains many components

• Wasteful to give whole blood if only red cells are needed


Blood Components

• Human blood consists of plasma, in which cells are suspended

• The plasma also contains other specialised substances, which are important for blood clot formation (e.g. clotting factors)

• Whole blood can be separated at the blood bank into various components


BLOOD COMPONENTS

• Blood separated into different parts:

1. Packed red cells2. Platelets3. Fresh frozen plasma4. Cryoprecipitate5. Granulocytes6. Factor IX conc.7. Factor VIII conc.

• There are more than 20 different products available


Whole blood

Red cells Plasma Platelets

(Fresh) frozen plasma (FFP)

CryoprecipitateStored Plasma

F lX

Immuno- globulins

Albumin

Fractionated products

F Vlla

F Vlll

Granulocytes


Whole Blood

• One unit contains • 450 ml of blood • & 63 ml of anticoagulant-preservative


RBC Anticoagulant/Preservative Solutions

• Purpose of RBC Preservation– Designed to prevent clotting and maintain red cell

viability and function during storage.– Usual anticoagulant-preservative is CPD-A (Citrate

Phosphate Dextrose Adenine )

• Anticoagulant-Preservative Contents• Citrate: anticoagulant (binds plasma calcium and prevent

activation of coagulation cascade)• Phosphate: provide substrate to help maintain red cell 2,3

DPG levels• Dextrose: a sugar, provides substrate for ATP production.• Adenine: Acts as a substrate for RBC synthesis of ATP


• During storage at 4oC

• Platelets and WBCs • become nonfunctional during hours of collection

• Red cells• 5 weeks in CPD-A have a mean recovery 70%

• Plasma K+, H+ → pH• Levels of coagulation factors V & VIII decrease


Blood Components

• Refers to a product separated from a single unit of whole blood

• The term plasma derivative indicates a blood product separated from a large volume of pooled plasma by a process called fractionation


• Blood components– Oxygen carrying components

– Red cell concentrates (RCC)

– Leukocyte poor blood– Frozen-thawed red cells

– Platelet products– Platelet rich plasma (PRP)– Platelet concentrates (PC)

– Plasma products– Fresh frozen plasma (FFP)– Frozen plasma (FP)– Cryoprecipitate– Stored plasma

• Plasma Derivatives– Coagulation Factor concentrates

• Factor VIII concentrates• Factor IX complex concentrates &

others

– Oncotic agents• Albumin• Plasma protein fraction (PPF)

– Immune serum Globulin• Hepatitis B Ig (HBIG)• Varicella-zoster Ig (VZIG)• Rh Ig (RhIG)• Tetanus Ig (TIG)


A- Blood components that carry oxygen

• Increase the oxygen carrying capacity of the blood by increasing the circulating red blood cell mass.

• Carry oxygen and nourishment to the tissues and take away carbon dioxide.


1- Red blood cell concentrates

• Prepared by removing approx. 200 ml of plasma from whole blood after centrifugation

• RBCs plus 100 ml of residual palsma

• In CPD-A can be stored for 35 days at 4oC



Whole Blood Red cell concentrate

Total Volume 500 ml 300 ml

Volume of red cells

200 ml 200 ml

Volume of plasma

300 ml 100 ml

Hematocrit 40 % 70 %



• High hematocrit → viscous → infuse slowly

• Rate of infusion increased by adding saline

• Other fluids should not used– Calcium containg fluids (eg. Ringer’s lactate)

should not be added– May cause clotting

– Glucose solutions – can cause clumping

• Only saline can be added to blood


2- Leukocyte poor blood

• No viable leukocytes• WBCs are of no

consequence • In some patients cause

febrile transfusion reaction• Should receive leukocytes

poor-blood• WBCs can be removed by

discarding the buffy coat (inverted centrifugation)

• Or by washing RBCs or by using filters

Buffy coat

Red cells


3- Frozen-thawed red cells

• Red cells can be frozen with use of cryopreservation techniques

• Permit storage for up to 10 years

• Expensive procedure & recommended only in special circumstances– e.g. Individuals with rare blood types– For auto-transfusion


3- Frozen-thawed red cells

• The RBC's are first incubated in a 40% glycerol solution which acts as an "antifreeze" within the cells.

• The units are then placed in special sterile containers in a deep freezer at less than -60 degrees C.

• Cryopreserved units are thawed and washed free of glycerol prior to use as saline suspended RBC's.


4- Synthetic oxygen carrying agents

• Synthetic oxygen carrying agents– Perfluorochemical (e.g. Fluosol-DA )

• Fluorinated hydrocarbons• Readily dissolve oxygen• Poor soluble in plasma• Side effects:

– Hypotension– DIC

– Chemically modified hemoglobin• Free Hb has a very short half life• Chemically modified to:

– increase intravascular survival – and to make it more effective in carrying oxygen


B- Platelet Products

• Platelet Rich Plasma (PRP)– Gentle centrifugation of whole blood– Supernatant transferred to the 2nd

bag• Platelet Concentrates

– Prepared from PRP by a 2nd centrifugation

– Removal of all but 50 ml of plasma– Contain approx. 6X1010 platelets– 60 – 80% Plts present in whole

blood unit– Remain 5 days– Longer at 22oC with continuous

agitation


B- Platelet Products

• Contamination by WBCs & RBCs is usually small

• But there is enough to induce alloimmunization

• Plt concentrates from Rh +ve should not be administered to Rh –ve women

• Storage at 22oC, therefore care to prevent contamination


C- Plasma Products

• Plt poor plasma can be separated into a number of products– Fresh frozen plasma– Frozen plasma– Cryoprecipitate– Stored plasma


1- Fresh frozen plasma (FFP)

• Prepared from whole blood within 6 hours of collection

• Rapid freezing of plasma preserves the labile coagulation factors at maximum levels

• Donot contain cellular elements

• 200 ml volume• Stored at -30oC for 12 months


2- Frozen Plasma (FP)

• Separated from whole blood within 24 hours of collection

• Contains at least 50 % of original factor VIII & factor V frozen plasma

• Adequate source for treatment of mild to moderate coagulation factor deficiencies

• 200 ml volume

• Storage at -30oC for up to 12 months


3- Cryoprecipitate

• Produced from freshly separated plasma by freezing at -70oC followed by thawing at 4oC

• Flocculent precipitate is rich in factor VIII, fibrinogen and fibronectin

• Once thawed, mixture is centrifuged to sediment the cryoprecipitate & all but 5 to 10 ml of supernatant plasma is removed

• Contains 250 mg fibrinogen• 80 clotting units of factor VIII• Stored at -30oC for 12 months


3- Cryoprecipitate

• Increase of 2% of factor VIII level for each bag of cryoprecipitate infused

• Supernatant plasma removed is called stored plasma– Must be used within 5 weeks if stored at 4oC– Lasts for 2 years at -30oC


4- Stored plasma

• Plasma separated from whole blood after 24 hours of storage at 4oC

• Can also be derived from cryoprecipitate production• Contain reduced levels of labile coagulation factors V VIII

& fibrinogen• It is indicated for patients requiring volume expansion or

protein replacement when labile clotting factors are not required

• Plasma products do not require crossmatch prior to use but should be ABO compatible


Plasma Derivatives


• Certain plasma derivatives can be obtained by fractionating the fresh frozen plasma or stored plasma

• Fractionation: Allows the processing of large volumes of

pooled plasma Pooling of many units increases the risk of

viral transmission to the recipient


Plasma protein fractionation

• Plasma proteins are separated according to differences of each protein.

• Fractionation involves changing the conditions of the pooled plasma (e.g. the temperature or the acidity)

• Proteins that are normally dissolved in the plasma fluid become insoluble, forming large clumps, called precipitate.

• The insoluble protein can be collected by centrifugation.


• One of the very effective ways for carrying out this process is the addition of alcohol to the plasma pool while simultaneously cooling the pool.

• This process is sometimes called cold alcohol fractionation or ethanol fractionation.

• This procedure is carried out in a series of steps so that a single pool of plasma yields several different protein products, such as albumin and immune globulin.


Plasma Derivatives

Plasma Derivatives Preparation avaliable

Coagulation Factors

Factor VIII concentrates

Factor IX concentrates

Anti-thrombin III

AlbuminAlbumin

Plasma protein fraction

Immune globulins

Non-specific immune serum globulin (ISG)

Rh immune globulin (RhIG)

Hepatitis B immune globulin (HBIG)

Varicella-Zoster immune globulin (VZIG)

Tetanus immune globulin (TIG)


1- Coagulation Factor Concentrates

• Prepared in a freeze-dried form

• Indicated for patients with congenital coagulation deficiencies– Risk of hepatitis is high

• Should not used for mild acquired coagulation deficiencies – Should be treated with FP or FFP


Factor VIII Concentrate

• Commercially prepared, lyophilized powder purified from human FFP

• Contain also small amounts of fibrinogen & other proteins

• Can contain blood group Abs • Treat patients with hemophilia A


Differences of Cryoprecipitate & Factor VIII concentrates

CryoprecipitateFactor VIII

concentrates

Storage Temp. -30oC4oC

Short period RT

Risk of Hepatitis

Low High

Treatment of hemophilia A

Yes Yes

Treatment of vW disease

Yes no


Factor IX Concentrate

• For the treatment Factor IX deficiency or Hemophilia B (Christmas Disease).

• Have been used to treat patients with acquired inhibitors of factor VIII– Have factor VIII bypassing activity

• Contains also factors II, VII & X in concentrated form

• Vials containing 500 units of factor IX


Factor IX Concentrate & liver disease

• It is contraindicated in patients with liver disease– Have low levels of circulating antithrombin III– Activation of clotting factors present in some

factor IX concentrates, – cause DIC


Blood products & treatment of specific clotting factor deficiencies

Deficiency Blood product Indicated

FibrinogenCryoprecipitate

Stored plasma

Factor VFresh frozen plasma

Frozen plasma

Factor VIIFactor IX concentrate

Stored plasma

Factor VIIIFactor VIII concentrate

Cryoprecipitate

Von Willebrand’s Disease

Cryoprecipitate

Fresh frozen plasma

Frozen plasma

Factor IX Factor IX concentrate

Factor X Stored plasma

Factor XI Stored plasma

Factor XIII Stored plasma


2- Oncotic Agents

• Albumin: volume expansion

• Other colloids are available for blood volume expansion– Dextran– Gelatin– Hydroxyethyl starch– Polyvinylpyrrolidone


Albumin

• Albumin is prepared by ethanol fractionation of pooled plasma

• Available in 5% and 25% concentrations.

• Have physiological sodium content

• No risk of hepatitis, sterilized during preparation

• No coagulation factors or blood group Abs


Albumin

• Used for treatment of hypovolaemia and hypoalbuminaemia (result from abnormal synthesis, increased metabolism or loss)

• It maintains capillary osmotic pressure

• Carrier protein for drugs, hormones, enzymes & metabolites


Plasma protein Fraction

• Partially purified albumin

• Contains ≈ 85% albumin & 15% other plasma proteins


3- Immune Globulins

• Contains immune IgG antibodies, prepared from pools of plasma.

• For disease prophylaxis, hepatitis A, measles, varicella and rubella.

• For the treatment of hypogammaglobulin-emia and agammaglobulinemia.


Immune Serum Globulin (ISG)

• Primarily IgG Ab

• Prevention of some viral diseases

• Hypogammaglobulinemia

• Congenital immune deficiency

• Given by IM injection (aggregates of IgG)


Hepatitis B Immune Globulin (HBIG)

• Contains Hepatitis B immune antibodies.

• From plasma of donors with high titer of Ab to HBsAg

• Provides passive immunization for HBV.

• For treatment after exposure to HBsAg.

• For the prevention of maternally transferred HBV (perinatal exposure).


Varicella-Zoster immune globulin (VZIG)

• Derived from patients had recent Herpes Zoster infections

• Herpes Zoster infections result in severe fatal infection in immunocompromised individuals

• Passive administration of VZIG during 72 hours of exposure can prevent or attenuate infection


Rh Immune Globulin (RhIG)

• Derived from Rh -ve individuals • Contains IgG antibodies to the D antigen on red

blood cells. • Given during pregnancy and post-natally to Rh

negative mothers to prevent the development of anti-D and hemolytic disease of the newborn (HDN) due to anti-D.

• Given prophylacticaly following abortion, or invasive maternal procedures (e.g., amniocentesis).


Tetanus Immune Globulin (TIG)

• Prepared from individuals specifically immunized for tetanus toxoid

• Available for individuals at risk following injury

Blood Components and Plasma derivatives

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