MUST TO KNOW IN IMMUNOHEMATOLOGY (BLOOD BANKING)

ISBT 001 ABO

ISBT 002 MNS
ISBT 003 P
ISBT 004 Rh
ISBT 005 Lutheran
ISBT 006 Kell
ISBT 007 Lewis
ISBT 008 Duffy
ISBT 009 Kidd
ISBT 010 Diego
ISBT 011 Cartwright
ISBT 012 Xg
ISBT 013 Scianna
ISBT 014 Dombrock
ISBT 015 Colton
ISBT 016 Landsteiner-Weiner
ISBT 017 Chido/Rodgers
ISBT 018 H
ISBT 019 Kx
ISBT 020 Gerbich
ISBT 021 Cromer
ISBT 022 Knops
ISBT 023 Indian
Chromosome 1 Rh
Duffy
Scianna
Cromer
Knops
Chromosome 2 Gerbich
Chromosome 4 MNS
Chromosome 6 Chido/Rodgers
Chromosome 7 Cartwright
Colton
Kell
Chromosome 9 ABO
Chromosome 11 Indian
Chromosome 17 Diego
Chromosome 18 Kidd
Chromosome 19 H
Lewis
Landsteiner-Weiner
Lutheran
Chromosome 22 P
Chromosome X Xg
Kx
Chromosome: Not known Dombrock
Von Descatello (Decastello) AB
Sturle (Sturli)
Blood groups (Most common) O > A > B > AB (Least common)

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Cell typing Forward/direct typing
Specimen: RBCs (Ag)
Reagents:
= Anti-A (blue: trypan blue)
= Anti-B (yellow: acroflavin dye)
= Anti-AB (colorless)
Uses of anti-AB Checks for the reaction of anti-A and anti-B
Detects weak subgroup of A and B because it has higher titer of anti-A & anti-B
Serum typing Reverse/indirect typing
Specimen: Serum (Ab) = 3-6 months (development)
Reagents:
= A1 cells
= B cells
Gel typing Major Advantage: Standardization
Medium: Dextran-acrylamide gel
0 = Unagglutinated Cells ---(media)---> Bottom of the μtube (microtube)
1+ = Agglutinated cells ---(media)---> May concentrate at the bottom of μtube
2+ = Agglutinated cells ---(media)---> Throughout the length of the μtube
3+ = Agglutinated cells ---(media)---> Concentrate near the top of the μtube
4+ = Agglutinated cells ---(media)---> Top of the media
Mixed-field:
= Agglutinated cells ---(media)---> Top
= Unagglutinated cells ---(media)---> Bottom
Red cell Ag-Ab reactions 0 = No agglutination/hemolysis
W+ = Tiny agglutinates, turbid BG
1+ = Small agglutinates, turbid BG (25% agglutination)
2+ = Medium agglutinates, clear BG (50% agglutination)
3+ = Large agglutinate, clear BG (75% agglutination)
4+ = One solid agglutinate (100% agglutination)
Universal donor Group O
Universal recipient Group AB
Universal donor for RBCs Group O (No Ag)
Universal recipient for Group AB (No Ab)
RBCs
Universal donor for plasma Group AB (No Ab)
Universal recipient for Group AB (No Ag)
plasma
RCS: Red cell suspension BB = 2-5% RCS
Approximate = Tomato red
Exact % RCS = [Vol. RBCs (mL) ÷ TV (mL)] x 100
Genotype One’s genetic makeup
Phenotype Expression of one’s genes
Homozygous In double dose
Heterozygous In single dose
Dominant Always expressed even in the heterozygous state
Recessive Not expressed when (+) dominant gene
To be expressed, it should be in the homozygous state
Allele One of two or more different genes that may occupy a specific locus on a
chromosome
Silent/amorph Only indicates the absence of the Ag
Von Dungern Subgroups of A = A1 (80%) and A2 (20%)
Hirzfeld
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Dolichos biflorus Lectin source of anti-A1
Phenotype Possible Genotypes
A1 A 1A 1
A 1A 2
A 1O
A2 A 2A 2
A 2O
A 1B A 1B
A 2B A 2B
B BB
BO
O OO
Gene Glycosyltransferase Immunodominant Sugar Ag Acceptor molecule
H L-fucosyltransferase L-fucose H Precursor subs.
A N-acetylgalactosaminyltransferase N-acetyl-D-galactosamine A H
B D-galactosyltransferase D-galactose B H
AB N-acetylgalactosaminyltransferase N-acetyl-D-galactosamine A H
D-galactosyltransferase D-galactose B
O -- -- -- Unchanged H
Amount of H Ag (Greatest) O > A2 > B > A2B > A1 > A1B (Least)
Bombay individual Bhende
(-) H gene
hh or Hnull
Lack H, A and B Ag’s
Designated as Oh
w/ anti-H, anti-A and anti-B Ab’s
Mistyped as group O
Parabombay Inherit weak H gene
w/ detectable A and B Ag’s but no detectable H Ag
Ah, Bh, ABh
Anti-H Differentiates Group O from Oh individuals
Ulex europaeus Lectin source of anti-H
ABO Histoblood group = present on all tissues and organs of the body
May be expressed in secretions depending on the secretor status (SeSe or Sese)
Determination of Secretor Specimen: Saliva
Status Principle: Hemagglutination-inhibition
ABO antibodies Mixture of IgM, IgG and IgA (Henry)
Anti-A Predominantly IgM
Anti-B React at room temp
Naturally occurring
Anti-A,B (Group O) Predominantly IgG
React at 37’C
Immune Ab
Immune Ab’s Production is stimulated by:
1. Pregnancy
2. Incompatible transfusion and transplant
ABO HDN Mother: Group O
Child: Group A or B
Group I discrepancies Weak reacting/missing Ab’s
Newborns
Elderly patients
Hypogammaglobulinemia/agammaglobulinemia
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Group II discrepancies Weak reacting/missing Ag’s (Less frequent)
Leukemia
Acquired B phenomenon
Subgroups of A
Hodgkin’s disease
BGSS (Remedy: Wash RBCs)
Group III discrepancies Plasma abnormalities resulting to Rouleaux formation (Plasma factor)
 globulins: MM, WM, HL
 fibrinogen
Dextran and PVP
Wharton’s jelly (Remedy: Wash cord cells 6-8x (7x)
Group IV Miscellaneous:
Polyagglutination
Anti-I (cold agglutinin)
Cis “AB phenotype”
Unexpected ABO isoagglutinins:
= Anti-H: produced by A1 and A1B (H Ag)
= Anti-A1: produced by A2 and A2B (No A1 Ag)
A Subgroups
A3 MF agglutination w/ anti-A & anti-A,B
Ax Weak MF agglutination w/ anti-A& anti-A,B
Aend MF agglutination w/ anti-A,B
Am (-) Agglutination, (+) AH substance in secretions
Ay (-) Agglutination,  AH substance in secretions
Ael (-) Agglutination, (+) H substance in secretions
B Subgroups
B3 MF agglutination w/ anti-B & anti-A,B
Bx MF agglutination w/ anti-A,B
Bm (-) Agglutination, (+) BH substance in secretions
Bel (-) Agglutination, (+) H substance in secretions
Rh Nomenclatures 1. Rosenfield = no genetic basis, indicates the presence or absence of Rh Ag’s
2. Fisher-Race (DCE) = inheritance of 3 genes (d= amorph/silent)
3. Wiener (Rh-hr) = inheritance of 1 gene (Ex. R0), w/c codes for an
agglutinogen (Ex. Rh0), w/c contains 3 blood factors (Ex. Rh0hr’hr’’)
Immunogenicity of Rh Ag’s (Most immunogenic) D > c > E > C > e (Least immunogenic)
R or r D or d
1 or ‘ Ce
2 or ‘’ cE
Z or y CE
G D+C+ red cell
Rh: 13, 14, 15, 16 4 different parts of the D mosaic
Genetic weak D D Ag’s expressed appear to be complete, but few in number
D Mosaic (Partial D) If one or more parts of D Ag is missing  weakened expression of D Ag
May produce anti-D (Ab against missing fragment)
4 fragments
C Trans D ---(trans)---> C (Ex. Dce/dCe)
f (ce) c ---(cis)---> e [Ex. Dce/DCE]
rhi (Ce) C ---(cis)---> e [Ex. DCe/DcE]
Hr0 “Common” Rh phenotypes (R1R1, R2R2, rr)
Rh Ab’s IgG1 and IgG3
React at 37’C
Immune Ab’s
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 (-) C’ binding = extravascular hemolysis (delayed HTR)
Rh HDN Mother: Rh (-)
Child: Rh (+), 2nd pregnancy
RhoGam or RhIg Purified anti-D
Administer w/in 72 hrs after 1st delivery
Full dose RhoGam 300 μg anti-D
(>12 weeks gestation) Protect up to 30mL D+ WB or 15mL D+ RBCs
Minidose/Microdose 50 μg anti-D
RhoGam Protect up to 5mL of D+ WB or 2.5mL D+ RBCs
(<12 weeks gestation) Ex. Abortion
# RhIg vials Volume of FMH (mL) ÷ 30
Vol. FMH = % fetal cells x 50
---------------------------------------------
x = (% Fetal cells x 50) ÷ 30
x ≈ __ + 1 = # RhIg vials
As little as 1mL Rh(+) RBC Produces anti-D
Rh+ RBCs + anti-D = (+) agglutination
Perform test for Du (IAT) If RBCs + anti-D = (-) agglutination
= IAT is (+) agglutination = +Du (weak D)
Rh- RBCs + anti-D + AHG reagent = (-) agglutination
2 conditions wherein an 1. No prior exposure to D Ag (males) or past childbearing age (females)
Rh- pt. can be transfused 2. Administer RhoGam
w/ Rh+ blood
Anti-LW Originally identified as anti-Rh in early experiments involving rabbits
(Landsteiner-Weiner) immunized w/ Rhesus monkey blood
Anti-LW agglutinates Rh+ and Rh- cells except Rhnull cells
Rhnull No Rh Ag
Designated as ---/---
Stomatocytes
Rhdeleted No C/c and E/e Ag
Designated as D--/D--
Lewis system Le gene codes for the production of fucosyltransferase enzyme that catalyzes
addition of fucose to the 4th C of N-acetylglucosamine of type 1 precursor
Lewis Ag’s Lea ---(Se)---> Leb
Produced by tissue cells
Not well developed at birth = NB/cord cells = Le(a-b-)
Decreased expression during pregnancy
Genotype Substances (Secretion) Phenotype Le Ab’s
ABH, lele, sese None ABH, Le(a-b-) Anti-Le & Anti-Leb
a

ABH, lele, SeSe/Sese ABH ABH, Le(a-b-) Anti-Lea & Anti-Leb

ABH, LeLe/Lele, sese Lea ABH, Le(a+b-) Anti-Leb
ABH, LeLe/Lele, SeSe/Sese ABH, Le , Le
a b ABH, Le(a-b+) none
Lewis Ab’s Anti-Le & Anti-Le
a b

Naturally occurring
IgM
Activates the C’
MN Ag’s Glycophorin A (MN-SGP)
M = Ser-Ser-Threo-Threo-Gly
N = Leu-Ser-Threo-Threo-Glu
Well developed at birth
Important in paternity testing
Anti-M IgM, pH-dependent (6.5), glucose-dependent
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Anti-N-like Ab IgM
Found in renal patients dialyzed w/ formaldehyde sterilized equipment
SS Ag’s Glycophorin B (Ss-SGP)
S = Methionine (29th)
s = Threonine (29th)
Ss Ab’s IgG
React at 37’C and AHG
Severe HTR w/ hemoglobinuria and HDN
Phenotype Detectable Ag’s Possible Ab’s
P1 P1 and P None
P2 P Anti-P1
p (p null) None Anti-P, P1, Pk (anti-Tja)
P 1k Pk and P1 Anti-P
P 2k Pk Anti-P, anti-P1
P1-like Plasma, pigeon and turtledove droppings, turtledove eggwhite
P1 substance Hydatid cyst fluid, Lumbricoides terrestris, Ascaris suum
Anti-P1 IgM
Naturally occurring
Strong anti-P1 = Hydatid disease (E. granulosus)
Associated w/ fascioliasis, C. sinensis and O. viverrini infections
Anti-Tja Anti-P, P1, Pk
Spontaneous abortions in early pregnancy
Anti-P IgG
Naturally occurring
Biphasic hemolysin (PCH)
Test: Donath-Landsteiner
Ii Blood Group I: Individuality
Neonates = I I (Ag) = I-i+
Adults (18 mos.-adult) = I I = I+i-
HEMPAS i Ag in adults
Anti-I Interfere w/ reverse typing (Group IV)
Benign anti-I = normal, IgM, naturally occurring, react at 4’C
Pathologic anti-I = IgM, react at 30/32’C (CAS = PAP)
Autoanti-I = M. pneumoniae, L. monocytogenes
Anti-i IgM
React at 4’C
EBV caused
Diseases of RES:
- Alcoholic cirrhosis
- Myeloid leukemia
- Reticuloses
Anti-IT Transition: from i  I
Yanomama Indians
Hodgkin’s lymphoma
K Kell
k Cellano
Kpa Penney
Kpb Rautenberg
Jsa Sutter
Jsb Matthews
Kell Ag’s Immunogenicity: 2nd to D (D>K>c>E>C>e)
Synthesized on precursor Kx
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 = On WBCs: remain unconverted. If (-)  CGD
= On RBCs: converted to Kell Ag’s. If (-)  MacLeod phenotype
MacLeod phenotype Acanthocytosis
Muscular dystrophy
Anti-K IgG
React at 37’C and AHG
Fy(a+b-) Chinese (90.8%)
Fy(a-b-) American blacks
Resistant to P. vivax/P. knowlesi (malaria)
Fya and Fyb Receptors for malaria
Duffy gene 1st human gene to be assigned to specific chromosome (1)
Anti-Fya IgG
Anti-Fyb React at AHG
Jka and Jkb Enhanced by enzymes
Anti-Jka Notorious antibody = not easily detected
Anti-Jkb Immune Ab’s (IgG)
Common cause of delayed HTR
Autoanti-Jk = Methyldopa (Aldomet)
Lua and Lub Development at age of 15
Anti-Lua IgG, IgM, IgA
Naturally occurring
React at room temp
Anti-Lub IgG4, IgM, IgA
Immune Ab
React at 37’C
Diego (Di) Mongolian ancestry
AE-1 = HCO3- <--> Cl-
Defect in AE molecule “ASO”
Acanthocytosis
Hereditary Spherocytosis
SEA Ovalocytosis
Cartwright (Yt) Erythrocyte acetylcholinesterase = neurotransmission
Xg Sex-linked
 Females
Short arm of X chromosome
Scianna Sc1, Sc2, Sc3
Dombrock Gregory (Gya)
Holley (Hy)
Joseph (Joa)
Colton (Co) CHIP, Aquaporin = water permeability
Chido/Rodgers (Ch/Rg) HLA system (Bg)
C4A (Rg) and C4B (Ch) = C’ component
HTLA = exhibit reaction only at high dilution
DAF (Cr)
Gerbich (Ge) GPC and GPD
Leach phenotype (GE: -2, -3, -4) = Elliptocytosis
Cromer (Cr) DAF
CROM Ab’s = black individuals
Knops (Kn) CR1 (C’ receptor 1)
Indian (In) CD44 (immune adhesion)
Benneth-Goodspeed (Bg) HLA Ag’s on RBCs (Class I MHC)
Bga = HLA-A7
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 Bgb = HLA-A17
Bgc = HLA-B28
Public Ag High-incidence Ag’s
Ena (99.9%)
Private Ag Low-incidence Ag’s (<1%)
HTLA Ab’s Anti-Ch
Anti-Rg
Anti-Kn
Anti-Yka
Anti-Csa
Anti-McC
Anti-JMH
Sources of Substances for Neutralization for Certain Antibodies
Anti-P1 Hydatid cyst fluid, pigeon droppings, turtledoves’ egg whites
Anti-Le Plasma/serum, secretor saliva
Anti-Ch, Anti-Rg Plasma/serum
Anti-Sda Urine, guinea pig urine
Anti-I Mother’s milk
Anti-H Saliva
Effect of Proteolytic Enzymes on Select Antigen-Antibody Reactions
Enhanced P1
Rh
I
Kidd
Lewis
Destroyed MNS
Duffy
Chido
Rodgers
Cartwright
Xg
Donation Process
At least 2 persons Bleeder
Head (BB)
Donor registration 1st step
Interview & physical exam 2nd step
Actual donor selection and 3rd step
blood collection
Collection <15 mins (7-10mins)
>15 mins = (-) cryoprecipitate
Donor bleeding 450 angle  10-200 angle
(Venipuncture: 150 angle)
Basic Qualifications of the Potential Blood Donor
Good health Purplish blue lesions  Kaposi’s sarcoma (HIV, HHV-8)
Age 18-65 yrs old
<18 yrs old = w/ parent’s consent
>65 yrs old = w/ physician’s consent
Body weight Max: 10.5mL/kg
Ideal: 110 lbs (50kg)
450mL blood + 30mL blood (serologic tests)
63mL anticoagulant

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Donor <110 lbs (<50kg):
= Volume of blood to be drawn
= Volume of anticoagulant
Vol. of blood to be drawn A = (Donor weight ÷ Ideal weight) x 450mL
Vol. of anticoag. needed B = (A ÷ 100) x 14
Vol. of anticoag. to be C = 63 – B
removed from blood bag
1:7 Anticoagulant: Blood ratio (Blood bag)
63mL AC = 450mL blood
31.5mL AC = 200mL blood
Temperature 37.5’C (99.5’F)
Pulse 50-100 beats/min ( athletes = acceptable)
BP cuff 40-60 mmHg (if used as tourniquet)
BP (DOH) 90-160 mmHg
60-100 mmHg
Min. H & H Allogeneic donation:
Hgb: ≥12.5 g/dL
Hct: ≥38%
Autologous donation:
Hgb: ≥11 g/dL
Hct: ≥33%
CuSO4 method Hgb determination
30mL container = 25 tests, solution changed daily
SG: CuSO4 = 1.053
1 cm = distance between blood and solution
Acceptable drop of blood  sink w/in 15 secs (Hgb: ≥12.5 g/dL)
Donor Deferral
Permanent Chagas’ disease
Babesiosis
Tegison (Tx: Psoriasis) = teratogenic
Recipient of human (pituitary)-derived GH = risk of transmitting CJD
[Recombinant GH = no deferral]
Recipient of cornea/dura mater = risk of transmitting CJD
3 years Malaria refugee/immigrant
1 year (12 months) Recipients of blood known to be possible sources of hepatitis
Tattoo
Rape
Incarceration in jail (3 days/72 hrs)
Blood transfusion
Major operation including dental surgery
Syphilis
Gonorrhea
Traveler  malaria-endemic places
Rabies vaccine
9 months (DOH) Childbirth (AABB: 6 weeks)
3 months (12 weeks) Recent blood donation (AABB: 8 weeks)
[DOH] = 450mL blood: 12 weeks
= 200mL blood: 6-8 weeks
1 month (4 weeks) Rubella vaccine
Isotretinoin/Accutane (Tx: Acne) = teratogenic
Proscar (Tx: Benign prostatic hyperplasia) = teratogenic
2-3 weeks After febrile episodes
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2 weeks Rubeola vaccine
Polio vaccine
Mumps vaccine
2 days (48 hrs) After hemapheresis
12 hrs After alcohol intake
Cause: Arterial puncture Jet-like pulsating bleeding w/ bright red blood
Citrate Binds Ca2+
Massive transfusion (8-10 units)  Citrate toxicity  Hypocalcemia
Milk = Ca2+
Dextrose/Glucose Provides energy for red cells
Citric acid Prevents caramelization (pH)
Adenine For improved survival of red cells
Phosphate buffer  ATP
Shelf-life: 21 days ACD (Apheresis)
CPD
CP2D
Shelf-life: 35 days CPDA-1
Shelf-life: 42 days CPDA-2 (DOH)
Additive solutions:
-Adsol (AS-1)
-Nutricel (AS-3)
-Optisol (AS-5)
Additive solutions SAGM:
-Saline
-Adenine
-Glucose
-Mannitol = RBC membrane stabilizing agent
Rejuvenation solutions  ATP & 2,3-DPG (Rejuvesol)
PIGPA = Phosphate, Inosine, Glucose, Pyruvate, Adenine
PIPA = Phosphate, Inosine, Pyruvate, Adenine
Heavy spin 5000g for 5mins = pRBC, platelet concentrate
5000g for 7mins = cryo, cell-free plasma
Light spin 2000g for 3mins = platelet-rich plasma
6-8 hours processing To maximize the number of components derived from 1 unit of blood
Platelet concentrate Light spin  Heavy spin
Centrifuge at 20-24’C
[Other blood components: Ref. centrifuge (1-6’C)]
Bacterial contamination
Separate from WB w/in 6-8 hrs
Blood Indication Storage Transport Shelf-life Other Info
Component
Whole blood 1. Blood vol. expansion &  1-6’C 1-10’C 21d (ACD, CPD) 1 unit:
RBC mass 35d (CPDA-1) Hgb by 1 to
2. Massive transfusion & 42d (CPDA-2) 1.5g/dL
acute blood loss 2d (Heparin Hct by 3-5%
Packed RBCs 1.  RBC mass 1-6’C 1-10’C Open system: 24h ‡80% plasma

(Normovolemic patients) Closed system: removed

21d (ACD, CPD) ‡(Hct: 65-80% but

35d (CPDA-1) not >80%)

1 unit:
Hgb by 1 to
1.5g/dL
Hct by 3-5%

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 Blood Indication Storage Transport Shelf-life Other Info
Component
Leukoreduced 1. FNHTR (Leuko. Ab’s) = 1-6’C 1-10’C Same as WB Preparation:
RBCs 1’C in temp. 1. Centrifugation
2. Prevent CMV 2. Filtration
3. Saline washing
Washed RBCs 1. Allergic (plasma w/ 1-6’C 24h Also for
foreign protein) polyagglutination
2. FNHTR (Leuko. Ab’s)
Anaphylactic: IgA w/ anti-
Iga
Frozen RBCs 1. Prolong storage of rare -65’C or 10 years
units -125’C
2. Prolong storage of
autologous units
Deglycerolized 24h
RBCs
Fresh frozen 1. Multiple coag. factor def. -18’C 1 year (-18’C) ‡Contains plasma,
plasma (liver disease) -65’C 7 years (-65’C) all coag. factors and
2. Replace isolated factor complement
def. when specific ‡FFP: Thaw at 37’C

component is not available ‡Thawed plasma:

3. Reverse effects of store at 1-6’C

coumarin/warfarin ‡Administer w/in

24hrs once thawed

Cryoprecipitate 1. Fibrinogen def. -18’C or 1 year ‡Contains:

2. Hemophilia A colder -150mg fibrinogen

3. vWD -80 U AHF
4. Factor XIII def. -vWF
-Factor XIII
‡Cryoppt.: Thaw at

37’C
‡Thawed cryoppt

(<15mL; not group-

spec.): store at RT’
‡Administer w/in 6

hrs
Granulocyte 1. Granulocyte dysfunction 20-24’C 24h Contains 1 x 1010
concentrate (CGD) w/o granulocytes
2. Myeloid hypoplasia agitation
Platelet 1. Thrombocytopenia 20-24’C 5d (20-24’C w/ 1 unit will 
concentrate w/ agitation) platelet count by
agitation 2d (1-6’C) 5,000- 10,000/μL
1-6’C
Irradiated blood 1. Prevent GVHD 28 days from Usually RBCs and
component irradiation or orig. platelets are
exp. date irradiated
whichever comes
first
Factor VIII 1. For hemophilia A 1-6’C Varies Stored in
concentrate lyophilized form
Factor IX 1. For hemophilia B 1-6’C Varies ‡A.k.a. prothrombin

concentrate complex
‡Contains factors II,

VII, IX and X
NSA 1. Hypovolemic shock 2-10’C 5 years 96% Albumin
4% Globulin

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 Blood Indication Storage Transport Shelf-life Other Info
Component
PPF 1. Hypovolemic shock 2-10’C 5 years 80-85% Albumin
10-15% Globulin
SDP 5 years SDP: Single donor
plasma
Methods of Freezing RBCs
Concentration Frozen at Stored at Equipment
High glycerol 40% w/v glycerol -80’C -65’C Mechanical
(Slow freezing) freezer
Low glycerol 20% w/v glycerol -196’C -120’C Liquid nitrogen
(Fast freezing)
Agglomeration Glycerol -80’C -65’C Mechanical
Glucose freezer
Fructose
EDTA
Cryoprotective agent Prevents rupture of RBCs during freezing
Ex. Glycerol
Deglycerolization Removal of glycerol
Hypertonic solution followed by an isotonic solution
High glycerol (DG) 12% NaCl -----> 1.6% NaCl -----> 0.9% NaCl
Low glycerol (DG) 45% NaCl in 15% mannitol
Agglomeration (DG) 50% Glucose + 5% Fructose -----> 0.9% NaCl
Cryoprecipitate After thawing = administer w/in 6 hours
After pooling = administer w/in 4 hours
Leukapheresis HES (Hydroxyethylstarch) = Separation bet. WBCs and RBCs
Donor: administered w/ corticosteroids 12-24 hrs before donation
= # of circulating granulocytes
Plateletpheresis Usually takes 1-2 hours
Donor:
= Platelet count: ≥150 x 109/L
= Aspirin-free: 3 days
Single donor platelets (SDP) A.k.a. super packed platelets
From plateletpheresis
For patients who are refractory or unresponsive to RDP
Limit patient exposure to multiple donors
RDP: Random Donor Platelets SDP: Single Donor Platelets
Preparation From WB: Light spin  Heavy spin Plateletpheresis
Amount 5.5 x 1010 platelets 3.0 x 1011 platelets
Vol. of plasma 50-75mL 300mL
pH ≥ 6.0 (New: 6.2) ≥6.0 (New: 6.2)
Storage 20-24’C w/ agitation 20-24’C w/ agitation
Shelf-life 5 days 5 days
At risk of TA-GVHD Recipient of BM transplant
Patients w/ hematologic or oncologic disease
Patients w/ congenital immunodeficiency
Recipient of blood from 1st degree relative (direct)
Irradiation Radiation source: 25-35 Gy (Gy: Gray | 1Gy = 100 rads)
a. Cesium (Ce)
b. Cobalt (Co)
Infusion IV fluids:
a. NSS = the only fluid allowed to start an IV line prior to transfusion

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 b. D5W (5% dextrose in H2O) = Not allowed to start (hypotonic  hemolysis)
c. Ringer’s lactate =not allowed to start (contains Ca2+  promote coagulation)
Blood warmers 37’C
Temp. >42’C = hemolyzed
Filters 1. Clot screen filter (170μm) = to remove gross clots
2. Leukocyte depletion filter
Speed of infusion 15gtts = 1mL
60gtts = 1min
1min = 4mL
1hr = 240mL
Rate = 240mL/hr
1 unit must be completed w/in 4hrs of blood transfusion
Polyagglutination (Hubener-Thomsen-Fridenrich Phenomenon)
Cryptantigens Hidden Ag’s
Covered w/ NANA (N-acetylneuraminic acid) or sialic acid
When NANA is destroyed (by neuraminidase)  Ag’s are exposed
Exposed Ag’s  Agglutination = React w/ tetrasaccharide of Thomas & Winzler
(T receptor)
Acquired Microbially-Associated Polyagglutination
T C. perfringens
V. cholerae
S. pneumoniae
All produce neuraminidase
Th E. coli
Proteus sp.
Produce weaker neuraminidase
Tx Bacterial and viral
Unknown mechanism
Tk “CABS”
C. albicans
A. niger
B. fragilis
S. marcescens
All produce endo/exogalactosidase
Altered precursor substances (Altered: ABH, Le, I, P)
Acquired B phenomenon Bacteria: Deacetylase enzyme
N-acetylgalactosamine --(Deacetylase)--> N-acetyl + galactosamine
Galactosamine = Group B
Remedy: Add acetic anhydride
VA Vienna, Virginia
Microbial fucosidase = fucose = H
Acquired Non-Microbially Associated Polyagglutination
Tn (-) β-3-D-galactosyltransferase enzyme needed for the normal structure of T
receptor (Tetrasaccharide of Thomas and Winzler)
Inherited Polyagglutination
Cad Sd a

HEMPAS/CDA II Adult i Ag = H Ag, Sialic acid

Others HbM – Hyde Park
NOR = Norfolk, Virginia

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 Tn Cad T Tk
Arachis hypogaea - - + +
Salvia sclarea + - - -
Salvia horminum + + - -
Glycine soja + + + -

Autologous Donation
Predeposit AD Before anticipated transfusion
Requirements:
*No age limit
*No strict weight requirement (vol. by 4mL/1 pound below 110)
*Hgb ≥ 11g/dL
*Hct ≥ 33%
*Frequency: every ≥ 3days
Intraoperative AD Collect blood during surgical procedure  reinfused immediately
Immediate preoperative Collect blood  replace patient volume w/ colloid/crystalloid  reinfuse
hemodilution during surgical procedure
Postoperative salvage Drainage tube
Postoperative bleeding  salvaged (saved)  clean and reinfused
Immediate Immune Transfusion Reactions
FNHTR 1’C in temperature
Cause: anti-leukocyte Ab’s (leukoagglutinins)
Prevention: Leukopoor RBCs
Allergic Cause: Donor plasma w/ foreign proteins
Prevention: Washed RBCs
Anaphylactic Afebrile (no fever)
Signs and symptoms occur only after the infusion of only few mL of blood
IgA deficiency w/ anti-IgA antibody
Prevention: Washed RBCs | IgA-deficient donor (rare)
Anaphylactoid Afebrile
Normal IgA w/ anti-IgA to donor IgA
Prevention: Washed RBCs | IgA-deficient donor (rare)
TRALI (NCPE) Cause: Anti-leukocyte Ab’s (leukoagglutinins)
Signs and symptoms resemble respiratory distress
Prevention: Leukopoor RBCs
Hemolytic Bleeding, hypotension, hemoglobinuria, anuria
Delayed Immune Transfusion Reactions
TA-GVHD Cause: T lymphocyte proliferation
Prevention: Irradiated RBCs
PTP Onset of thrombocytopenia
Cause: anti-platelet Ab’s (HPA-1a negative platelets)
Prevention: Therapy
a. Administration of corticosteroid
b. Exchange transfusion
c. IV immunoglobulins
DHTR 7 days
Immediate Nonimmune Transfusion Reactions
TACO Administration of blood w/o equivalent blood loss
Iatrogenic: physician-caused
At risk:
a. Young children
b. Elderly patients
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 c. Patients w/ cardiac disease
Prevention: Therapy
a. Therapeutic phlebotomy
b. IV diuretics
c. O2 therapy
Bacterial contamination Cause: Endotoxin production by psychrophilic/cryophilic bacteria
Y. enterocolitica (most common)
E. coli
P. aeruginosa
Factors:
a. During phlebotomy
b. During preparation/processing
c. During thawing
Prevention:
a. Sterile technique
b. Visual inspection of unit
→ Blood unit = Brown, purple, hemolysis, clot
→ Plasma = Murky (dark brown) purple, red
PCITR Causes:
- Small bore needle
- Warming blood above 50’C
- Freezing blood w/o cryoprotective agent
- Citrate toxicity
Delayed Nonimmune Transfusion Reactions
Iron Overload Patients w/ normovolemic anemia
(Hemosiderosis) Transfusion-dependend patients:
- Aplastic anemia
- Congenital hemolytic anemia
Prevention:
a. Iron-chelating agent = Deferroxamine
b. Neocytes = young RBCs, has longer lifespan
Disease transmission HBV, HCV, HDV, CMV, EBV, HTLV-I and II, HIV, T. pallidum, Plasmodium spp., B.
microti, T. cruzi, T. gondii
Hemolytic Disease of the Newborn
In utero Anemia ( immature RBCs, enlarged spleen & liver = extramedullary
hematopoiesis)
Hydrops fetalis = cardiac insufficiency, edema
Neonatal period  Unconjugated bilirubin  Brain  Kernicterus
Treatment 1. Intrauterine transfusion
- In utero
- Corrects anemia
- X-match: Mother’s serum
2. Exchange transfusion
- Neonatal period
- Removes bilirubin & Ab-coated RBCs
- X-match: Mother’s serum (preferred) or infant’s serum
Cross-Matching
Full X-match 2 hours
Can be shortened to 30 mins
Abbreviated X-match Type/screen + immediate spin
DC/PS = no agglutination/hemolysis
Electronic X-match Patient blood type is determined on 2 occasions

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Emergency situation Patient blood type is unknown
Group O (Rh-) pRBCs
(+) Major X-match (-) Alloantibody control
(+) Autoantibody, autocontrol
Ratio of serum to cells Routine = 40:1
Det. weak Ab’s = 133:1
Saline for washing pH 7.2-7.4
Incubation period 30-120 mins (majority: 30 mins)
Enhancement media 1. Albumin
2. PEG
3. LISS (Incubation pd: 5-15 mins)
Lectin Sources
Anti-A1 Dolichos biflorus
Anti-H Ulex europaeus
Anti-M Iberis amara
Iberis umbellate
Iberis semperivens
Maclura aurantiaca
Anti-N Bauhinia variegate
Bauhinia purpura
Bauhinia bonatiana
Bauhinia candicans
Vicia graminea
Anti-B Bandeirae simplicifolia
Quality Control
Annually Mercury thermometers
Quarterly (Every 3 months) Speed timer (centrifuge)
Cell washers (speed, timer)
Blood warmers
Monthly Alarm Activation (refrigeration and freezers)
Temperature (refrigerated centrifuge)
Daily Refrigerators and freezers (continuous monitors)
Platelet incubators (enclosed, monitored chambers)
Daily when in use Transfusion service:
- Heating blocks
- Water baths
Donor facility:
- Donor unit agitators
- Scales
- Balances
- Hemoglobinometer
- Microhematocrit centrifuges
Every 4 hours Platelet incubators (ambient temperature storage)
Other Topics
Apheresis/Hemapheresis Whole blood is withdrawn, a desired component separated, and the remainder
of the tube returned to the donor
Intermittent-flow 1 venipuncture
centrifugation (IFC) Blood is withdrawn and reinfused through the same needle
Once the desired component is separated, the remaining components are
reinfused to the donor
Continuous-flow 2 venipunctures
centrifugation (CFC) Withdraw, process and return the blood to the individual simultaneously
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Antibody screen
Donor & recipient samples
Compatibility testing
DHTRs
Virus inactivation in plasma
products
Y. enterocolitica
Density
Requirements for blood
products to be transfused
to infants
Allogeneic donor blood
from outside sources
w/in 30 mins
Procedure when HTR
occurs
Specimens for investigation
of HTR
LISS
ZZAP
True chimerism
Artificial chimerism
AB cis genotype
Acquired A antigen
Collect new specimen every 3 days in a series of transfusion
Keep for 7 days after transfusion (at 1-6’C)
Series of tests that ensure safety of transfusion to recipient and donor
1. Review of BB records
2. ABO and Rh
3. Ab screen
4. Cross matching
a. Major X-match = DC/PS
b. Minor X-match = PC/DS
Occur 3 to 7 days after transfusion
Heating in a liquid state w/ LMW stabilizers
Heating in lyophilized state
UV irradiation
 Bacterial contamination of blood
Separates neocytes from mature RBCs
1. (-) HbS
2. <7 days
3. γ-irradiated
4. (-) CMV
Repeat ABO and Rh typing
Time limit when a unit of blood is removed from 2-8’C and returned back to the
refrigerator
1. Stop transfusion
2. Keep IV line open
3. Notify the physician and BB
1. Patient pre-transfusion blood sample
2. Patient post-transfusion blood sample
= PT/PTT/DAT
= Pink: Hgb 0.2 g/L or 20 mg/dL
= Red: Hgb >1 g/L or 100 mg/dL
3. Patient post-transfusion urine
= Bilirubin/Urobilinogen
4. Blood bag
= GS/CS
Glycine/glucose + saline
5-15 mins incubation
Cysteine-activated papain
Mixture of DTT and papain
Presence of 2 cell population
Ex. Twins
Mixed-field agglutination
After:
-BM transplantation
-Blood transfusion
-Exchange transfusion
-Fetomaternal hemorrhage
Group IV discrepancy
Inheritance of 3 ABO genes (AB/O)
1. Tn activation
2. P. mirabilis
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Acquired B antigen “EPIC”
1. E. coli O86
2. P. vulgaris
3. Intestinal obstruction
4. Carcinoma of the colon and rectum
HAT medium Hypoxanthine, Aminopterin, Thymidine medium
Culture for hybridoma cells
Hybridoma cells Plasma cells (mouse) + myeloma cells + PEG
Polyspecific AHG reagent Anti-IgG and anti-C3d
Prepared by conventional technique (rabbits)
Monospecific AHG reagent Anti-IgG or anti-C3d
Prepared by monoclonal Ab production (mice)
Type 1 precursor substance β 1 3
ABH substance in secretions (glycoproteins)
HAB, Se, Le genes
Type 2 precursor substance β 14
ABH on RBC (glycoproteins)
HAB genes
Agglutination in Gel Incompatible
technology
Affinity column technology Affinity adherence of IgG-sensitized RBCs to immunologically active matrix
(Gamma ReACT)
Wash RBCs 2-3x to remove unbound globulins
# cryobags [Desired level of Factor VIII x volume of plasma (mL)] ÷ 80
# units to test # units required ÷ frequency of donors (-) for relevant Ag
Enzyme technique 1. Papain = papaya
2. Bromelin = pineapple
3. Ficin = figs
4. Trypsin = pig’s stomach
Elution Breaking of bonds between Ag and Ab by:
a. Physical: heating, shaking
b. Chemical: ether, acid
Adsorption Removal of Ab in serum using appropriate RBC
Biphasic hemolysin Cold = attaches to RBCs
Warm = RBC lysis
Perfluorocarbons O2 carrying capacity
RBC substitute

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