blood banking

1. B L O O D
C O M P O N E N T S A N D
P R E P A R A T I O N
Dr. Amany M. Elshamy
Lecturer of Biochemistry and
Molecular Diagnostics
PH.D of Biochemistry and
Molecular biology
MLS, AUC, Cairo

2. O U T L I N E S

3. I N T RO D U C T I O N
The concept is, when a single unit of whole blood can benefit many
patients, why waste it on one patient and transfuse him the
components, which he does not require at all.
The introduction of integrated plastic blood bags and the refri-
gerated centrifuge in the blood banking has completely
revolutionised the transfusion therapy.
Hemapheresis is a process that uses an apheresis machine to
selectively remove components from donor blood.

4. O V E R V I E W O F T H E R O U T I N E B L O O D B A N K L A B O R A T O R Y
• Blood and blood components must be stored and shipped
under conditions that ensure their viability.
• Every donor unit is tested to determine ABO group and Rh
type, including weak D when indicated. Units are also tested
for selected viral markers.
• The type of Rh-negative units must be confirmed because of
potential sensitization that may occur if Rh-positive blood is
transfused to an Rh-negative recipient.

5. • The following components can be separated from a single unit of
450ml blood:
• Red cell concentrate (Packed red cells)
• Platelet concentrates
• Fresh frozen plasma (FFP)
• Cryoprecipitate
• Leucocyte-depleted RBC concentrate
• Granulocyte concentrate
• Single donor plasma
• Fibrinogen concentrate
• Factor VIII concentrate.

7. P R O C E S S I N G
O F W H O L E
B LO O D U N I T S
F R O M
C O L L E C T I O N
TO L A B E L I N G

8. RIA, ELISA (or EIA), WB,
and IF techniques are
immunologic
methods based on
detection and
quantification of
antigen or antibody by
the use of a
radioisotope, enzyme,
or fluorescent labels

10. P R E PA R AT I O N O F R B C C O N C E N T R AT E
The RBC concentrate has a hematocrit of almost 80-95%.
The following procedure is applied for the preparation of RBC
concentrate.
To prevent graft versus host disease, blood components prepared for a
fetus that needs an intrauterine transfusion should be irradiated.

11. P R E PA R AT I O N O F R B C S C O N C E N T R AT E
Centrifugation method
• Collect blood preferably in a 450 ml
of double or triple bag system.
• The blood bags are kept tightly
packed and balanced in the buckets of
the refrigerated centrifuge.
• Centrifuge at 5000 g for 5 minutes at
4-6° C.
• Express almost 3/4th of plasma in
the satellite bag.
• Seal the tube in between,
connecting the two bags.
• Detach the satellite bag
• Keep the bag with red cells at 4-6° C
• Store the plasma bag at – 30° C
• The shelf life of the RBC concentrate
is the same as that of whole blood,
depending on the preservative used.

13. S T A N D A R D T E S T S
• Leukocyte-reduced blood has been
filtered to remove the white blood
cells that contain antibodies that can
cause fevers in the recipient of the
transfusion. (These antibodies, with
repeated transfusions, may also
increase a recipient's risk of reactions
to subsequent transfusions.)

14. L E U C O C Y T E S D E P L E T E D R E D C E L L
C O N C E N T R AT E
• Leucocyte induced non-
haemolytic febrile reactions
can be minimized by reducing
the number of leucocytes to
0.5 × 109 in the RBC
concentrate.

15. L E U C O C Y T E S D E P L E T E D R E D C E L L
C O N C E N T R AT E
The methods for depletion of leucocytes are as follows:
• Removal of buffy coat by centrifugation.
• Washing of red cells
• Spin and filtration
• Freezing and deglycerolisation.

16. L E U C O C Y T E
D E P L E T I O N B Y
C E N T R I F U G AT I O N
• The procedure is simple and reliable.

19. L E U C O C Y T E
D E P L E T I O N B Y
C E N T R I F U G A T I O N
• This could be achieved by
two methods: Upright
Spin and Inverted spin

20. L E U C O C Y T E D E P L E T I O N B Y C E N T R I F U G AT I O N
Upright Spin
• Collect blood in 450 ml blood bags.
• Keep the bags in the bucket of the centrifuge and balance
properly.
• Centrifuge at 5000 g for 5 minutes at 4-6° C.
• Express the plasma, buffy coat and approximately 10-20 ml of
upper layer of red cells in the satellite bag.
• Seal the connecting tube and separate the bags.

21. S T A N D A R D T E S T S
• Irradiation to blood cells is performed
to disable any T-lymphocytes present
in the donated blood. (T-lymphocytes
can cause a reaction when transfused,
but can also cause graft-versus-
host problems with repeated exposure
to foreign cells.)

22. P R E PA R AT I O N O F F R E S H F R O Z E N P L A S M A
( F F P )
• The plasma collected from a single donor or by plasmapheresis and
frozen within 4 hours at –70°C is called fresh frozen plasma (FFP).
• Procedure
• Collect 450 ml blood in a double or triple bag.
• The collected blood must be stored at 4-6°C for not more than 4 hours
prior to centrifugation.

23. P R E PA R AT I O N O F F R E S H F R O Z E N P L A S M A
( F F P )
• Centrifuge the bags at 5000 gm for 5 minutes at 4-6°C in a properly
packed and balanced position.
• Express 3/4th of the plasma in the satellite bag.
• Seal the tube and detach the plasma-containing satellite bag.
• Freeze the plasma at -70° C in a mechanical freezer within 4 hours.
• The ideal volume of FFP is 200- 250 ml.
• Store the plasma in an inverted position at –30°C (stored for 1 year).

24. P R E PA R AT I O N O F P L AT E L E T C O N C E N T R AT E
( P C )
• Like FFP the platelet concentrate can also be prepared from single
donor unit or by apheresis.
• Procedure
• Collect blood in a 450 ml triple or quadruple bag, after having a
clean neat venepuncture.
• There should be a free flow of blood preferably collected within 8
minutes of venepuncture.
• Place the bags in the buckets and balance them properly.
• First centrifuge at 2000 g for 3 minutes (light spin) at 20-24°C to
produce platelet-rich plasma (PRP).

25. P R E PA R AT I O N O F P L AT E L E T C O N C E N T R AT E
( P C )
Procedure:
• Express the PRP in the platelet bag and seal the tube between the
primary bag and two satellite bags.
• Centrifuge again at 5000 g (heavy spin) for 5 minutes at 20-24°C.
• Express the plasma into the second satellite bag leaving the platelet
concentrate in the platelet bag.
• The ideal volume of platelet concentrate is 40-70 ml.
• Store the platelet concentrate at RT (20-24°C) in a shaker or
agitator for 5 days only

26. P L AT E L E T C O N C E N T R AT E ( P C )
• The specifications for an ideal platelet concentrate are as
follows (quality control):
• Volume – 40-70 ml
• Platelet yield – >5.5 × 1010
• Leucocytes – < 108
• RBC – < 0.5 ml
• pH – > 6.0

27. N O T E ! !
• The donor who has taken Aspirin in the last 3 days
must not be accepted as a donor for PC.
• The platelets become haemostatically ineffective in
the presence of circulating salicylic acid in the blood.
• The acetylation of platelets cyclooxygenase leads to
reduced production of thromboxane A2.

28. C R Y O P R E C I P I TAT E
• This component consists of precipitated plasma proteins,
fibrinogen, factor VIII, and von Willebrand factor.
• The cryo is prepared by rapid freezing within 6 hours and
then subsequent thawing of the FFP at 1-6°C.
• The standard cryo bag should contain at least 80 IU of factor
VIII.

29. C R Y O P R E C I P I TAT E
Preparation of cryoprecipitate
• Collect 450 ml of blood in a triple integrated bag and prepare FFP.
• Express the plasma into the satellite bag and clamp the tubing between
the primary and satellite bag.
• Freeze the plasma, and keep the RBC at 2-6°C.
• Thaw the plasma at 4°C in a shaking water bath or in a refrigerator at 2-
6°C.

30. C R Y O P R E C I P I TAT E
Preparation of cryoprecipitate
• Centrifuge both bags using heavy spin to separate cryoprecipitate from
plasma. Open the closure between the primary and satellite bag.
• Express the plasma back into the red cells in the primary container.
• Seal the tubing twice and cut between the seals to separate the two
bags.
• Immediately refreeze the cryoprecipitate.
• Keep the whole blood at 2-6°C in the refrigerator.
• Cryoprecipitate that has been pooled must be transfused within 4 hours.

32. Q S

33. • Which of the following statements is true concerning human
blood?
• a)The blood of all normal humans contains red and white cells,
platelets, and plasma.
• b)Some human populations normally lack the ability to produce
plasma.
• c)Proteins are not normal components of human blood.
• What is the shipping temperature requirement for plasma?
• a. 1°F to 6°F or higher
• b. 1°C to 6°C or lower
• c. _18°F or higher
• d. _18°C or lower

34. • Which of the following are likely to increase in quantities when the body
is under attack from bacteria?
• a)erythrocytes
• b)leukocytes
• c)thrombocytes
• When blood clumps or forms visible islands in the still liquid plasma, it is
called:
• a)clotting
• b)agglutination
• c)none of the above

35. • Antigens are:
• a)found on the surface of red cells
• b)kinds of red cells that identify a blood type
• c)relatively large carbohydrate molecules
• d)a and b
• Most of the volume of normal human blood is composed
of:
• a)red cells
• b)hemoglobin
• c)plasma
• d)white cells

36. • Whole blood collected from a donor should remain at what temperature before the
separation in preparation of platelet concentrates?
• To prevent graft versus host disease, blood components prepared for a fetus that
needs an intrauterine transfusion should be----------------
• Rh factor derives its name from which of the given options?
(a) Ape
(b) Human
(c) Rat
(d) Monkey

37. • Give the 3 main components made from donated whole blood
• Red Cells Human
• Insulin
• Plasma (FFP / Cryo)
• Platelets
• Granulocytes
• Anti-D
• Normal Saline
• Growth hormone

38. • Which of the following anticoagulant preservatives provides a storage time of 35 days at 1°C
to 6°C for units of whole blood and prepared RBCs if an additive solution is not added?
• a. ACD-A
• b. CP2D
• c. CPD
• d. CPDA-1
• Additive solutions are approved for storage of red blood cells for how many days?
• a. 21
• b. 42
• c. 35
• d. 7

39. • What are the current storage time and storage temperature for platelet concentrates
and apheresis platelet components?
A 5 days at 1C-6C
B 5 days at 24C-27C
C 5 days at 20c-24C
D 7 days at 22C-24C
• Whole blood and RBC units are stored at what temperature?
A 1C to 6C
B 20C to 24C
C 37*C
D 24C to 27C

40. • One criterion used by the FDA for approval of new preservation solutions and storage
containers is an average 24-hour post-transfusion RBC survival of more than:
A 50%
B 60%
C 65%
D 75%
• Frozen and thawed RBCs processed in an open system can be stored for how many
days/hours?
A 3 days
B 6 hours
C 24 hours
D 15 days
•

41. • Which of the following occurs during during storage of red
blood cells?
A pH decreases
B 2,3-DPG increases
C ATP increases
D Plasma K+ decreases
• Nucleic acid amplification testing is used to test donor blood
for which of the following diseases?
A Hepatitis C virus
B HIV
C West Nile virus
D all of the above