COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

PLATELET CONCENTRATES
Can be produced during the routine conversion of whole blood into concentrated rbcs or by apheresis essential for the formation of the primary hemostatic plug and maintenance of normal hemostasis FOR : Actively bleeding patients (thrombocytopenic, less than 50 x 109/L due to decreased production or decreased function) Cancer patients during radiation and chemotherapy because of induced thrombocytopenia ( less than 20 x 109/L) Thrombocytopenia preoperative patients (less than 50 x 109/L)

PLATELET CONCENTRATES
Random-donor platelets
Prepared from whole blood Should contain at least 5.5 x 1010 Stored at 20 24OC with continuous agitation Contain sufficient plasma to yield a pH of greater than or equal to 6.2 Shelf life 5 days

Single-donor platelets
prepared from apheresis Contain at least 3.0 x 1011 Stored at 22 24OC with agitation Contain approx. 300 ml of plasma Shelf-life 5 days

Not usually indicated for patients with DIC or idiopathic thrombocytopenic purpura (ITP)

PLATELET CONCENTRATES
Single-donor platelets
Generally indicated for patients who are unresponsive to random platelets due to HLA alloimmunization To limit the platelet exposure from multiple donors

PLATELET CONCENTRATES
Platelet concentrate preparation prepared within 6 hours of collection
Leave approx. 50 70 ml of plasma on the platelets To maintain a pH of 6.2 during storage Plasma stored as FFP, single-donor plasma frozen within 24 hours (PF24) or liquid recovered plasma Platelet concentrate lie undisturbed for 1 to 2 hours at 20 24OC, making sure the platelet button is covered with plasma Shelf-life is 5 days from the date of collection If open system, transfusion must occur within 6 hours Label properly (volume, expiration date and time) All units for a single dose ( 6 to 8 units) must be pooled and must be transfused within 4 hours of pooling

Platelet concentrate preparation prepared within 6 hours of collection

Centrifuge whole blood (set at 22OC) light spin 3200 rpm for 2 to 3 minutes Platelet rich plasma is expressed into one of the satellite bags Disconnect rbc and store it at 4OC Recentrifuge PRP at 22OC using a heavy spin 3600 rpm for 5 minutes Express majority of the plasma into the second satellite bag

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

PLATELET CONCENTRATES
QUALITY CONTROL PROCEDURES (monthly)
Platelet count (random-donor 5.5 x 1010 and single-donor 3.0 x 1011) Volume must be sufficient to maintain an acceptable pH until the end of the dating period methods in place to limit and detect bacterial contamination in platelet components
improved phlebotomy techniques culture and staining methods to detect bacterial organisms dipsticks to detect bacterial levels a swirling technique to detect metabolic changes in platelets

PLATELET ALIQUOTS
Indicated for neonates whose counts fall below 50,000/mL and who are experiencing bleeding Factors that may be associated with thrombocytopenia:
1. 2. 3. 4. 5. 6. immaturity of the coagulation system platelet dysfunction increased destruction of platelets dilution effect secondary to massive transfusion or exchange Transfusion intraventricular hemorrhage

Either random or apheresed platelets may be transfused

Expiration: 4 hours

Can be irradiated, leukocyte-reduced or washed

PLATELETS LEUKOREDUCED
PURPOSE OF LEUKOREDUCTION:
1. Prevention of febrile nonhemolytic reactions 2. Prevent or delay the development of HLA antibodies 3. Reduce the risk of transmission of CMV

GRANULOCYTES PHERESIS
For Patients who have received:
intensive chemotherapy for leukemia bone marrow transplant, OR BOTH

must contain less than 8.3 x 105 leukocytes If the platelets have been pooled, a method must be used that results in a leukocyte count of less than 5 x106 in the final pooled product
Single-donor or apheresis platelets that have been leukoreduced must contain less than 5 x 106 leukocytes in at least 95% of units tested

CRITERIA:
those with fever, neutrophil counts less than 500/mL septicemia or bacterial infection unresponsive to antibiotics reversible bone marrow hypoplasia a reasonable chance for survival

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

GRANULOCYTES PHERESIS
Adults: usual dose is1 granulocyte pheresis product daily for 4 or more days For neonates, a portion of a granulocyte pheresis unit is usually given once or twice Granulocytes components should be administered as soon as possible and within 24 hours of collection

PLASMA
Separated from cellular blood elements and frozen to preserve the activity of labile coagulation factors Plasma must be placed in the freezer within time frame required for the anticoagulant or collection process
Stored at 18OC

Cryoprecipitation is accomplished by slow thawing, at 1 to 6OC, plasma that has been prepared for freezing within the time frame required for the anticoagulant or collection process
Cryoprecipitate should be refrozen within 1 hour of thawing Store at 18 C or colder, preferably 30 C or colder, for up to 12 months from the date of blood collection

SINGLE-DONOR PLASMA: FFP

Frozen plasma from single donors may comprise FFP, PF24, or plasma cryoprecipitate-reduced Fresh frozen plasma (FFP)
produced from whole blood when obtained from a single, uninterrupted nontraumatic venipuncture plasma is frozen within 8 hours of collection (CPD, CD2D, or CPDA-1 within 6 hours if the preservative was ACD stored at 18OC for 1 year or 65OC for 7 years contains maximum levels of labile and nonlabile clotting factors about 1 IU per ml

SINGLE-DONOR PLASMA
Plasma frozen within 24 hours after phlebotomy (PF24/FP24)
frozen within 8 to 24 hours of collection Stored at 18OC contains all stable proteins found in FFP

Both PF24 and FFP are thawed at temperatures between 30 and 37OC or in an FDA-approved microwave device
Product should be stored within 1 to 6OC for no more than 24 hours (after thawing) Indicated in patients who are actively bleeding and have multiple clotting factor deficiencies

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

SINGLE-DONOR PLASMA
A single unit of FFP or PF24:
should contain 150 to 250 mL of plasma approximately 400 mg of fibrinogen 1 unit of activity per mL of each of the stable clotting factors

PLASMA AND LIQUID PLASMA

Can be prepared directly from whole blood or as a by-product of platelet concentrate or cryoprecipitate production Can be used as volume expanders or for the manufacturing of plasma fractionation products such as:
plasma protein fraction (PPF) normal serum albumin (NSA) immune serum globulin (ISG) Plasma in a unit of whole blood can be separated during storage and up to 5 days after the expiration date Stored at 18OC or colder and labeled as plasma with a shelf-life of 5 years can be transfused up to 5 days after the expiration date of the whole blood

FFP also contains the same level (1 unit/mL) of factors V and VIII Plasma cryoprecipitate reduced (cryopoor plasma)
Supernatant remaining from the production of cryoprecipitate

used exclusively in the treatment of thrombotic thrombocytopenic purpura.

Cryoprecipitate has been removed from the plasma product and stored at 18OC or colder, expiration is 1 year from date of collection

S/D POOLED PLASMA

Consists of pools of no more than 2500 units of ABO typespecific plasma treated with solvent/detergent in the thawing process to inactivate lipid-enveloped viruses such HIV and hepatitis B Solvent tri-n-butyl phosphate Detergent triton X-100
Product volume is approximately 200 mL Contains stable and labile clotting factors but lacks von Willebrands factor multimers Does not protect against the nonlipid enveloped viruses such as hepatitis A and B19 parvovirus

CRYOPRECIPITATED ANTIHEMOPHILIC FACTOR

Cold-precipitated concentration of factor VIII, the antihemophilic factor (AHF) Prepared from FFP thawed slowly (14 16 h)between 1 and 6OC Suspended in less than 15 mL of plasma Contains: most of the factor VIII and part of the fibrinogen from the original plasma at least 80 units of AHF activity and at least 150 mg of fibrinogen Others: factor XIII and von Willebrands factor has a shelf-life of 12 months in the frozen state and must be transfused within 6 hours of thawing or within 4 hours of pooling

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

CRYOPRECIPITATED ANTIHEMOPHILIC FACTOR

Like FFP and PF24, cryoprecipitate should be thawed quickly at 37OC Once thawed, store at room temperature (22 to 24OC until transfused Indicated in the treatment of:
1. 2. 3. 4. Classic hemophilia (hemophilia A) von Willebrands disease Factor XIII deficiency and as a source of fibrinogen for hypofibrinogenemia

CRYOPRECIPITATE
Cold insoluble portion of plasma remaining after FFP has been thawed at refrigerator temperatures

Constituents Factor VIII Fibrinogen vWF Factor XIII

Amount 80 120 U/concentrate 150 250 mg/concentrate 40 70% of original FFP 20 30% of original FFP

Store at 18C or colder up to 12 months from the date of whole blood collection

NOVOSEVEN
Recombinant activated factor VII (NovoSeven, Denmark) Induces hemostasis in life- and limb-threatening bleeds and in major surgery of hemophilia A and B patients in the presence of inhibitors Factor VIIa binds to activated platelets and activates small amounts of FX independent of tissue factor and that the platelet surface FXa can restore platelet surface thrombin generation in hemophilia useful in hemophilia patients who have developed inhibitors to factor VIII

FACTOR VIII CONCENTRATES

Used in the treatment of classic hemophilia and hemophilia A and in persons deficient in factor VIII Prepared from large volumes of pooled plasma or in the form of recombinant FVIII using DNA technology Techniques used to inactivate or eliminate viral contamination:
1. Pasteurization 2. Solvent/detergent treatment 3. Monoclonal purification

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

FACTOR VIII CONCENTRATES

1. Pasteurization
Stabilizers such as albumin, sucrose, or glycine are added to the factor VIII concentrate to prevent denaturation of the product Heated to 60OC for 10 hours Stabilizers are removed, and the product is lyophilized

PORCINE FACTOR VIII

Xenographic form of factor VIII Made from porcine plasma Beneficial for patients with hemophilia A who have developed inhibitors or antibodies to human factor VIII Shown to provide effective hemostatic control for patients with intermediate FVIII inhibitor levels Residual porcine vWF in the preparation of the product induces platelet activation
Enhancing hemostasis apart from the action of circulating FVIII

2. Solvent/detergent treatment
Use of ethyl ether and tri(n-butyl) phosphate and the detergent sodium cholate and Tween 80 Removed then lyophilized

3. Monoclonal purification
Immunoaffinity chromatography
used to positively select out of pooled plasma the vWF:FVIII complex

RECOMBINANT FACTOR VIII

Production of recombinant human FVIII (rFVIII) Synthesized by introducing human FVIII gene into BHK (baby hamster kidney cells) rFVIII released into culture medium and harvested, isolated, and purified using a combination of ion exchange chromatography, gel filtration, and immunoaffinity chromatography Purification and final formulation Human albumin as a stabilizer Next generation : rFVIII:FS uses sucrose as final stabilizer

FACTOR IX CONCENTRATES
Three forms: 1. Prothrombin complex concentrates
contains significant levels of vitamin Kdependent factors: II, VII, IX, and X prepared from large volumes of pooled plasma by absorbing the factors out using barium sulfate or aluminum hydroxide lyophilized and virally inactivated may contain activated vitamin Kdependent factors

2. Factor IX concentrates
developed by monoclonal antibody purification less thrombogenic than prothrombin complex concentrates

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

FACTOR IX CONCENTRATES
2. Factor IX concentrates
contains approximately 20 to 30% of FIX stored in the refrigerator in lyophilized form

FACTOR XIII CONCENTRATES

Factor XIII deficiency a severe autosomal-recessive bleeding disorder associated with a characteristic pattern of neonatal hemorrhage and a lifelong bleeding diathesis There are currently two plasma-derived virus inactivated factor XIII concentrates: 1. Fibrogammin P
available in Europe, South America, South Africa, and Japan

3. Recombinant factor IX (rFIX)

produced in a Chinese hamster ovary cell line and not thought to transmit human infectious disease treatment of patients with hemophilia B

2. Factor XIII concentrate (Bio Products Laboratory, Elstree, UK)

UK

IMMUNE SERUM GLOBULIN

Immune globulin is indicated for:
1. Patients with immunodeficiency diseases (i.e. severe combined immunodeficiency and Wiskott-Aldrich syndrome) 2. Providing passive antibody prophylaxis against hepatitis and herpes

IMMUNE SERUM GLOBULIN

IVIg is also used in patients with:
Idiopathic thrombocytopenic purpura Post-transfusion purpura HIV-related thrombocytopenia Neonatal alloimmune thrombocytopenia (NAIT)

Concentrate of plasma gamma globulins in an aqueous solution Prepared from pooled plasma by cold ethanol fractionation Administered IV or IM Half-life of 18 to 32 days

Contraindications:
Individuals with a history of IgA deficiency or anaphylactic reactions should not receive immune globulin
because of the presence of trace amounts of IgA

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

NORMAL SERUM ALBUMIN (NSA)

Prepared from salvaged plasma Pooled and fractionated by a cold alcohol process Treated with heat inactivation (60OC for 10 hours), which removes the risk of hepatitis or HIV infection Composed of: 96% albumin and 4%globulin Albumin used routinely as the replacement fluid in many plasmapheresis procedures
replacing the colloid that is removed during the procedures

NSA
Available in 25% or 5% solutions Intended for:
patients who are hypovolemic hypoproteinemic In clinical settings of shock burn patients

25%preparation is contraindicated in patients who are dehydrated

unless it is followed with crystalloid infusions (e.g., normal saline) for volume expansion

Can be used with diuretics to induce diuresis in patients with low total protein because of severe liver or protein-losing disease

PLASMA PROTEIN FRACTION (PPF)

Contains 83% albumin and 17% globulins Similar in preparation with NSA with lesser purification steps available in a 5% preparation Similar use to that of NSA contraindicated for infusion during cardiopulmonary bypass procedures PPF and NSA can be stored for 5 years at 2 to 10 OC and have not been reported to transmit HIV or hepatitis

RhO (D) IMMUNOGLOBULIN

Rh immunoglobulin (RhIg) is a solution of concentrated antiRho(D) prepared from pooled human plasma of patients who have been hyperimmunized and contains predominantly IgG antiD Has 2 primary uses: (1) treatment of ITP and (2) prevention of Rh HDN During the first 12 weeks of pregnancy, a 50-mg dose of RhIg is indicated for D-negative females for abortion or miscarriage

/Maida Fatima Diaz Estira-Chan

COMPONENT PREPARATION & TRANSFUSION THERAPY

3/8/2013

RhO (D) IMMUNOGLOBULIN

After 12 weeks gestation, a full dose (300 mg) is indicated for abortion or miscarriage in D negative women 120-mg dose is advised after 34 weeks gestation:
When amniocentesis is performed In the event of obstetric complication or following termination of pregnancy

SYNTHETIC VOLUME EXPANDERS

TWO CATEGORIES: 1. Crystalloids
Ringers lactate consists of Na, Cl, K, Ca, and lactate ions normal isotonic saline Na and Cl ions

2. Colloids
Dextran prepared in a 6 and 10% solution with a half-life of 6 hours HES available in a 6% solution with an IV half-life of more than 24 hours

RhIg is also used in the event Rh-positive components are transfused to Rh-negative patients
A 300-g dose IM (120-mg dose IV) is sufficient to protect against Dpositive RBCs contained in 10 units of random platelets

used as volume expanders in hemorrhagic shock and burn patients

SYNTHETIC VOLUME EXPANDERS

CHARACTERISTIC Intravascular Retention Peripheral edema Pulmonary edema Easily excreted Allergic reactions Cost Examples COLLOID Good Possible Possible No Rare Expensive Albumin Dextran Hydroxyethyl starch CRYSTALLOID Poor Common Possible Yes Absent Inexpensive Ringers lactate solution 7.5% Normal saline

ANTITHROMBIN III CONCENTRATES

Antithrombin-III concentrates (AT-III) or antithrombin (AT) an inhibitor of clotting factors IX, X, XI, XII, and thrombin prepared from pooled human plasma and heat-treated to prevent viral transmission for treatment of patients with hereditary AT deficiency in connection with surgical or obstetrical procedures Or when they suffer from thromboembolism rhAT is produced by transgenic goats expressing recombinant human AT in their milk

/Maida Fatima Diaz Estira-Chan