Blood Components and Plasma derivatives M Zaharna Blood

Blood Components and Plasma derivatives M. Zaharna Blood Bank Lab. 2009 1

• 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 M. Zaharna Blood Bank Lab. 2009 2

Blood collection • Blood is collected in plastic bag systems with anticoagulant & preservative • Whole blood can be stored at 4 o. C for up to 5 weeks • Whole blood contains many components • Wasteful to give whole blood if only red cells are needed M. Zaharna Blood Bank Lab. 2009 3

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 M. Zaharna Blood Bank Lab. 2009 4

BLOOD COMPONENTS • Blood separated into different parts: 1. 2. 3. 4. 5. 6. 7. • Packed red cells Platelets Fresh frozen plasma Cryoprecipitate Granulocytes Factor IX conc. Factor VIII conc. There are more than 20 different products available M. Zaharna Blood Bank Lab. 2009 5

Whole blood Red cells Granulocytes Platelets Plasma Fractionated products (Fresh) frozen plasma (FFP) F Vlla Cryoprecipitate Stored Plasma Immunoglobulins F Vlll Albumin F l. X M. Zaharna Blood Bank Lab. 2009 6

Whole Blood • One unit contains • 450 ml of blood • & 63 ml of anticoagulant-preservative M. Zaharna Blood Bank Lab. 2009 7

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 M. Zaharna Blood Bank Lab. 2009 8

• During storage at 4 o. C • Platelets and WBCs • become nonfunctional during hours of collection • Red cells • 5 weeks in CPD-A have a mean recovery 70% • Plasma • K+, H+ → p. H • Levels of coagulation factors V & VIII decrease M. Zaharna Blood Bank Lab. 2009 9

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 M. Zaharna Blood Bank Lab. 2009 10

• Blood components • Plasma Derivatives – 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 – – – Coagulation Factor concentrates • Factor VIII concentrates • Factor IX complex concentrates & others – Oncotic agents • Albumin • Plasma protein fraction (PPF) – Immune serum Globulin Fresh frozen plasma (FFP) Frozen plasma (FP) Cryoprecipitate Stored plasma M. Zaharna Blood Bank Lab. 2009 • • Hepatitis B Ig (HBIG) Varicella-zoster Ig (VZIG) Rh Ig (Rh. IG) Tetanus Ig (TIG) 11

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. M. Zaharna Blood Bank Lab. 2009 12

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 4 o. C M. Zaharna Blood Bank Lab. 2009 13

1 - Red blood cell concentrates Whole Blood Total Volume 500 ml Red cell concentrate 300 ml Volume of red cells Volume of plasma Hematocrit 200 ml 300 ml 100 ml 40 % 70 % M. Zaharna Blood Bank Lab. 2009 14

1 - Red blood cell concentrates • 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 M. Zaharna Blood Bank Lab. 2009 15

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 M. Zaharna Blood Bank Lab. 2009 Buffy coat Red cells 16

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 M. Zaharna Blood Bank Lab. 2009 17

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. M. Zaharna Blood Bank Lab. 2009 18

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 M. Zaharna Blood Bank Lab. 2009 19

B- Platelet Products • Platelet Rich Plasma (PRP) – Gentle centrifugation of whole blood – Supernatant transferred to the 2 nd bag • Platelet Concentrates – Prepared from PRP by a 2 nd centrifugation – Removal of all but 50 ml of plasma – Contain approx. 6 X 1010 platelets – 60 – 80% Plts present in whole blood unit – Remain 5 days – Longer at 22 o. C with continuous agitation M. Zaharna Blood Bank Lab. 2009 20

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 22 o. C, therefore care to prevent contamination M. Zaharna Blood Bank Lab. 2009 21

C- Plasma Products • Plt poor plasma can be separated into a number of products – Fresh frozen plasma – Frozen plasma – Cryoprecipitate – Stored plasma M. Zaharna Blood Bank Lab. 2009 22

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 -30 o. C for 12 months M. Zaharna Blood Bank Lab. 2009 23

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 -30 o. C for up to 12 months M. Zaharna Blood Bank Lab. 2009 24

3 - Cryoprecipitate • Produced from freshly separated plasma by freezing at -70 o. C followed by thawing at 4 o. C • 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 -30 o. C for 12 months M. Zaharna Blood Bank Lab. 2009 25

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 4 o. C – Lasts for 2 years at -30 o. C M. Zaharna Blood Bank Lab. 2009 26

4 - Stored plasma • Plasma separated from whole blood after 24 hours of storage at 4 o. C • 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 M. Zaharna Blood Bank Lab. 2009 27

Plasma Derivatives M. Zaharna Blood Bank Lab. 2009 28

• 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 M. Zaharna Blood Bank Lab. 2009 29

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. M. Zaharna Blood Bank Lab. 2009 30

• 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. M. Zaharna Blood Bank Lab. 2009 31

Plasma Derivatives Preparation avaliable Factor VIII concentrates Coagulation Factors Factor IX concentrates Anti-thrombin III Albumin Plasma protein fraction Non-specific immune serum globulin (ISG) Rh immune globulin (Rh. IG) Immune globulins Hepatitis B immune globulin (HBIG) Varicella-Zoster immune globulin (VZIG) Tetanus immune globulin (TIG) M. Zaharna Blood Bank Lab. 2009 32

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 M. Zaharna Blood Bank Lab. 2009 33

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 M. Zaharna Blood Bank Lab. 2009 34

Differences of Cryoprecipitate & Factor VIII concentrates Factor VIII Cryoprecipitate concentrates o. C 4 Storage Temp. -30 o. C Short period RT Risk of Low High Hepatitis Treatment of Yes hemophilia A Treatment of Yes no v. W disease M. Zaharna Blood Bank Lab. 2009 35

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 M. Zaharna Blood Bank Lab. 2009 36

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 M. Zaharna Blood Bank Lab. 2009 37

Blood products & treatment of specific clotting factor deficiencies Deficiency Blood product Indicated Cryoprecipitate Fibrinogen Stored plasma Fresh frozen plasma Factor V Frozen plasma Factor IX concentrate Factor VII Stored plasma Factor VIII concentrate Factor VIII Cryoprecipitate Von Willebrand’s Disease Fresh frozen plasma Factor IX concentrate Factor X Stored plasma Factor XIII Stored plasma M. Zaharna Blood Bank Lab. 2009 38

2 - Oncotic Agents • Albumin: volume expansion • Other colloids are available for blood volume expansion – Dextran – Gelatin – Hydroxyethyl starch – Polyvinylpyrrolidone M. Zaharna Blood Bank Lab. 2009 39

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 M. Zaharna Blood Bank Lab. 2009 40

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 M. Zaharna Blood Bank Lab. 2009 41

Plasma protein Fraction • Partially purified albumin • Contains ≈ 85% albumin & 15% other plasma proteins M. Zaharna Blood Bank Lab. 2009 42

3 - Immune Globulins • Contains immune Ig. G antibodies, prepared from pools of plasma. • For disease prophylaxis, hepatitis A, measles, varicella and rubella. • For the treatment of hypogammaglobulinemia and agammaglobulinemia. M. Zaharna Blood Bank Lab. 2009 43

Immune Serum Globulin (ISG) • • • Primarily Ig. G Ab Prevention of some viral diseases Hypogammaglobulinemia Congenital immune deficiency Given by IM injection (aggregates of Ig. G) M. Zaharna Blood Bank Lab. 2009 44

Hepatitis B Immune Globulin (HBIG) • Contains Hepatitis B immune antibodies. • From plasma of donors with high titer of Ab to HBs. Ag • Provides passive immunization for HBV. • For treatment after exposure to HBs. Ag. • For the prevention of maternally transferred HBV (perinatal exposure). M. Zaharna Blood Bank Lab. 2009 45

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 M. Zaharna Blood Bank Lab. 2009 46

Rh Immune Globulin (Rh. IG) • Derived from Rh -ve individuals • Contains Ig. G 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). M. Zaharna Blood Bank Lab. 2009 47

Tetanus Immune Globulin (TIG) • Prepared from individuals specifically immunized for tetanus toxoid • Available for individuals at risk following injury M. Zaharna Blood Bank Lab. 2009 48