0312 Antibody Id

Antibody Identification
Part 1
Part 1: The Basics
Prerequisites
Geography of a panel
Antibody ID method
Case examples
A Blood Bank Guy Video Podcast
D. Joe Chaffin, MD
March 2012
www.bbguy.org
Tuesday, March 6, 12
Prerequisites
When Do We I.D.?
Blood Group Overview

General facts
Podcast from December 2011
Pretransfusion Testing
Testing methodologies
Antibody screen
Podcast from February 2012
Following a positive antibody screen

Patients, prenatals, donors
When testing suggests a new antibody
To confirm a previously identified
antibody (per facility SOP)
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3
Definitions
Whats a Panel?
Alloantibody
Antibody against RBC antigens
not present on patients own RBCs
Autoantibody
Antibody against RBC antigens
Just an expanded antibody screen

Uses group O reagent RBCs
RBCs from 8-20 donors
Patient serum or plasma
IS / 37 C / AHG if tubes
AHG only if gel or solid phase
Reactions documented on a sheet
present on patients own RBCs
that outlines every RBCs phenotype
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Geography
Lets take a close look at a panel
Important to know your way around
There are variations, but this is a
general guide
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A series of 8-20 group O

donor reagent red cells,
each tested for all main
antigens (phenotyped)
A series of 8-20 group O

donor reagent red cells,
each tested for all main
antigens (phenotyped)
Rh-hr (or Hr) =

Modified Wiener Rh
genotype for donor
Rh-hr (or Hr) =

Modified Wiener Rh
genotype for donor
R1: DCe
R2: DcE
R0: Dce
RZ: DCE
r: dCE
r: dcE
r: dce
ry: dCE
R1: DCe
R2: DcE
R0: Dce
RZ: DCE
Other stuff:
-Donor ID
-Special Antigen Type
r: dCE
r: dcE
r: dce
ry: dCE
Other stuff:
-Donor ID
-Special Antigen Type
Co(b+)
Pt. phenotype results
+
D, C, c, E, e, Fya, Fyb, Jka, Jkb,

K, k, Lea, Leb, M, N, P1, S, s
+
10
Jk(a+b): Assume double dose Jka

Jk(a+b+): Single dose Jka and Jkb
Jk(ab+): Assume double dose Jkb
Full donor phenotype

D phenotype for each donor,
C phenotype for each donor,etc...
*
*
*
*
*
*
*
*
11
12
Jka
Jka
Cell 1
Jka
Jkb
FyaCell 2
Fya
Jkb
Jkb
IS 37 AHG
0
0
0
0
0
0
0
0
0
0
0
a
FyCell
5
Fy
13
IS AHG
0
0
0
0
0
0
0
0
0
0
3+
3+
3+
3+
0
0
0
0
0
0
This is tube testing!

LISS
Albumin
Saline
PEG
IS = Immediate spin
37 = 37 C Incubation
AHG = Anti-human globulin
IAT = Indirect antiglobulin test
(=AHG)
14
AHG
3+
3+
3+
3+
0
0
0
0
0
0
This is also tube testing!

(often with PEG)
0 3+
1+ 3+
1+ 3+
1+ 3+
1+ 3+
0 0
0 0
0 0
0 0
0 0
0 0
1+ 3+
IS = Immediate spin
(=AHG)
IS = Immediate spin
(=AHG)
This could be:

-Tube testing
OR
-Gel testing
OR
-Solid-phase testing
3+
15
16
Pos Control
Neg Control
3+
0
0
0
0
0
0
3+
4+
0
IS = Immediate spin
(=AHG)
3+
3+
3+
3+
0
0
0
0
0
0
3+
This could be:

-Tube testing
OR
-Gel testing
OR
-Solid-phase testing
17
3+
3+
0
0
0
0
0
0
3+
0
IS = Immediate spin
(=AHG)
This is almost always:

-Gel testing
OR
-Solid-phase
18
Ya Gotta Have a Plan...
All other results

are suspect
Consistent approach minimizes error

Most are simple, but cutting corners
increases risk for dumb mistakes
Use this or your own system, but use
If the AC is positive:
the same approach every time!
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General Process
History
Both in real life and on exams
History can give you a clue and keep
you from doing something dumb
Up to 70% of cases impacted by
Check history
Check autocontrol
Look at general pattern
Look at whats NOT there (cross-outs)
Look at what IS there
Use special techniques as necessary
Ensure statistical significance
history
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History
History
Clinical history examples:

Anti-D in pregnant pt; consider RhIG
Recent bacterial infection; consider
antibiotic induced warm autoab
Recent viral illness; consider autoanti-I or -i (consider age)
Recent transfusion; consider newly
developing antibody
ITP; consider IV RhIG in D+ patient
Clinical history examples:

Consider racial profiling (in a good
way, of course!)
African-Americans: Lack of Duffy
antigens
Asians: Almost all D+
Whites: May lack high freq antigens
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History
General Process
Consider serologic history, if known

Previous phenotyping will help, but
be careful!
Transplants, transfusions, errors
In real world, can use this info for
targeting the panel and cells chosen
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Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
k Kpa Kpb Jsa Jsb Fya Fyb Jka Jkb P1 Lua Lub M N
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
26
Positive Autocontrol
Results
Lewis
s Lea Leb Cell
AC
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Rh-hr
Cell Rh-hr D C
Check history
Check autocontrol
More details next time...

Question 1: Is DAT positive?
Question 2: Whats the patient history?
Possibilities include:
Autoantibodies (warm and cold)
Recent transfusion/DHTR
Drug-induced issues
Passively acquired antibodies
3+
0
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28
General Process
Pattern
Are reactions:
Uniform or variable?
Against all, most, or rare cells?
Present in what phases?
Check history
Check autocontrol
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Variability
Cells Reactive
Uniform reactions suggest a single

antibody
Variable reactions suggest either:
Multiple antibodies OR
Single antibody with dosage
With negative autocontrol...

A mixture of reactive and
nonreactive cells suggests:
A single alloantibody OR
Multiple alloantibodies
A single reactive cell suggests:
A single alloantibody against a
low-prevalence antigen
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37/AHG: Rh
Cells Reactive
AHG:
K, Fy, Jk, S/s
Variable: Lu, Xg
IS (RT):
With negative autocontrol...

All or virtually all cells reactive
suggests:
Multiple alloantibodies
A single alloantibody against a
37 C and AHG V
Le, MN, P
AHG Only
RT Var. RT AHG RT
high-prevalence antigen
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AC neg
AC neg
Variable
Uniform
Warm
All +
1. IgG allo vs high prev
2. Multiple IgG allos
3. HTLA-like
Warm
Cold
+&
All +
+&
1. Single IgG allo

1. IgM allo vs high prev
1. Single IgM allo

2. Multiple IgM allos
Many/most +
2. Single IgG allo (dosage)
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Warm/Cold
Many/most +
1. IgG/IgM allos (multiple)
2. Strong IgM binding
1. Allo vs high prev

antigen (IgG or IgM)
All +
Cold
Many/most +
1. Multiple IgM allos
2. Single IgM allo (dosage)
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36
Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
General Process
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11
R1R1
+ + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11 3+
AC
AC
Check history
Check autocontrol
1. Gel or solid-phase most likely (probably gel)

2. No autoantibody obvious
3. Uniform pattern; single antibody most likely
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Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Cross-outs
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
AC
3+
0

38
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Cross-outs
Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11
R1R1
+ + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11 3+
AC
AC
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Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
42
Results
Lewis
Rh-hr
s Lea Leb Cell
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
AC
3+
0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
43
AC
3+
0
44
Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Results
Lewis
Rh-hr
s Lea Leb Cell
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11
R1R1
+ + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11 3+
11
R1R1
+ + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11 3+
AC
AC
AC
AC

45
General Process
General Process
Try first for a single antibody to
Check history
Check autocontrol
explain all reactions
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DC E Cw V Kpa Jsa Lua

Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
General Process
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11
R1R1
+ + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11 3+
AC

Failing that...
Depending on pattern, hypothesize:
Two antibodies in same phase
AC

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Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 4+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 4+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 2+
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
General Process
Results
Lewis
s Lea Leb Cell
AC

Failing that...
One warm and one cold antibody
4+
0

3. Variable pattern; multiple antibodies most likely
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Fyb
Colds: Lea, Leb, M, N, P

Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Lewis
s Lea Leb Cell
Results
IS 37 IAT
0 0 0
R1R1
+ + 0 0 + 0 + 0 +
0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0+ +0 0 + 0 + 2 0
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + +
0 0 0 3 0
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 0
rr
rr
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7
rr
9
10
Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Lewis
s Lea Leb Cell
Results
IS 37 IAT
0 0 0
R1R1
+ + 0 0 + 0 + 0 +
0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1
0 0
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0+ +0 0 + 0 + 2 0
0 0
0 0
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + +
0 0 0 3 0
0 0
0 0
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 0
0 0
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 1+ 1+ 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 1+ 1+ 3+
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 1+ 1+ 1+
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 1+ 1+ 1+
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 +
0 +
0 + 0 + + + + 0 + 0 8
0 0 3+
1+ 1+ 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 +
0 +
0 + 0 + + + + 0 + 0 8 1+ 1+ 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9
0 0 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 1+ 1+ 1+
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 1+ 1+ 1+
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
AC
0 0 3+
0 0 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0
+ + 0 0 + + 0 0 + 0 + 11
AC
1. Tube testing for sure

3. Highly variable pattern, both warm and cold
53
Rh-hr
c
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Lewis
s Lea Leb Cell
54
R1R1
+ + 0 0 + 0 + 0 +
0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0+ +0 0 + 0 + 2 0
0 0
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + +
0 0 0 3 0
0 0
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 0
0 0
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 1+ 1+ 3+
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 1+ 1+ 1+
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 +
0 +
0 + 0 + + + + 0 + 0 8 1+ 1+ 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 1+ 1+ 1+
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0
+ + 0 0 + + 0 0 + 0 + 11
AC
AC
General Process
Results
IS 37 IAT
0 0 0

Failing that...
One warm and one cold antibody
Consider multiple warms and colds
Next podcast!!
0 0 3+
0 0 0
0 0 3+
0 0 0

0 0 3+
0 0 0
Fyb , Lea
Colds: Lea, Leb, M, N, P

E
0 0 0

Cell Rh-hr D C
AC
0 0 3+
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56
General Process
Phenotyping
Helps confirm identification of
Check history
Check autocontrol
alloantibody by demonstrating lack

of antigen
Is a TOOL in confirmation, not sole
measure of confirmation
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57
58

Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Adsorption
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 4+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 4+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 2+
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
Removing antibodies from sample by
incubation with antigen-positive RBCs

Autoadsorption
Patient
K Patient
K
Autoantibody
4+
AC 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
0 0
Anti-K

3. Variable pattern; multiple antibodies most likely
p. 7
59
AutoAb
Patient
K
AutoAb
Patient
K
Anti-K
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60
Adsorption
Elution
Removing antibodies from sample by
Removal of RBC-bound antibodies

Heat, cold, chemical (glycine)
incubation with antigen-positive RBCs

Alloadsorption
Anti-KK+C+S
Anti-CK+C+S
Anti-SK+C+S
p. 7
Elution
A
-K ntiC
nti
K Anti
ti-C
An
K+C+S
-K
nti
An
ti-
A K+C+S C
A K+C+S
K Anti
ti-
An K+C+S C
Anti-S
Adsorbed Serum
p. 7
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61
Treat
Anti-C
K+C+S
or...
K+C+S
www.bbguy.org
Proteolytic Enzymes
62
General Process
Enzymes such as ficin and papain may
change Ag expression/Ab binding
M M M M
M
M
M
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Anti-K
Check history
Check autocontrol
www.bbguy.org
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64
Probability
Probability
Basic idea: Ensure what you are

seeing is not pure chance
Traditional interpretation:
Ag present: 3 positive reactions
Ag absent: 3 negative reactions
AABB Standards for IRLs, 7th ed:
5.3.3 requires only 2 of each
For practical purposes, preliminary

ID only requires ONE double-dose
rule out to establish lack of identity
This does NOT mean your work is
done, however
Still need to use selected cells to
rule in/out specific Abs (per OP)
reaction to assign specificity
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65

Rh-hr
Cell Rh-hr D C
Kell
f Cw V
Duffy Kidd P
Lutheran
MNSs
Lets do some
together!
Results
Lewis
s Lea Leb Cell
R1R1
+ + 0 0 + 0 + 0 0 + 0 + 0 + + 0 + 0 + 0 + 0 + 0 + 0 + 1 3+
R1R1
+ + 0 0 + 0 0 0 0 + 0 + 0 + + 0 + + 0 0 + 0 + 0 + 0 + 2 3+
R2R2
+ 0 + + 0 0 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + + + 0 0 3 3+
R0r
+ 0 0 + + + 0 0 0 + 0 + 0 + 0 0 + 0 + 0 + 0 + 0 + 0 0 4 3+
rr
0 + 0 + + + 0 0 0 + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 + 0 5 0
rr
0 0 + + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + 0 + + + + 0 6 0
rr
0 0 0 + + + 0 0 + + 0 + 0 + 0 + 0 + + 0 + + 0 + 0 0 + 7 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + + + 0 + 0 + + + + 0 + 0 8 0
rr
0 0 0 + + + 0 0 0 + 0 + 0 + + 0 + + + 0 + + + + + 0 + 9 0
10
rr
0 0 0 + + + 0 0 + + 0 + 0 + + + + + + 0 + + 0 0 + + 0 10 0
11 R1R1 + + 0 0 + 0 0 0 + + 0 + 0 + 0 + 0 + 0 0 + + 0 0 + 0 + 11
AC
AC
3+
0

66
www.bbguy.org
67
68
Thanks!
This podcast is not a medical consultation. Please
consult your provider before making any medical
decisions, and do not use the information presented
here for anything other than educational purposes.
Monica LaSarre (Bonfils)

Tuan Le (Bonfils)
Colleen Chiappa (Bonfils)
Kevin Elman (N. Co. Med Center)
Cami Melland (Bonfils)
The immortal Connie Howard
Please do not reproduce or rebroadcast

this podcast without my permission!
Seriously! Youre getting it for free, right?
(Walter Reed)
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www.bbguy.org
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70

0312 Antibody Id

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0312 Antibody Id

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Antibody Identification

Part 1: The Basics

A Blood Bank Guy Video Podcast

Blood Group Overview

Following a positive antibody screen

Just an expanded antibody screen

present on patients own RBCs

that outlines every RBCs phenotype

A series of 8-20 group O

A series of 8-20 group O

Rh-hr (or Hr) =

Rh-hr (or Hr) =

Pt. phenotype results

D, C, c, E, e, Fya, Fyb, Jka, Jkb,

Jk(a+b): Assume double dose Jka

Full donor phenotype

This is tube testing!

This is also tube testing!

This could be:

This could be:

This is almost always:

Ya Gotta Have a Plan...

All other results

Consistent approach minimizes error

the same approach every time!

Clinical history examples:

Clinical history examples:

Consider serologic history, if known

targeting the panel and cells chosen

More details next time...

Uniform reactions suggest a single

With negative autocontrol...

K, Fy, Jk, S/s

With negative autocontrol...

1. Single IgG allo

1. IgM allo vs high prev

1. Single IgM allo

1. Allo vs high prev

1. Gel or solid-phase most likely (probably gel)

1. Gel or solid-phase most likely (probably gel)

s Lea Leb Cell

s Lea Leb Cell

1. Gel or solid-phase most likely (probably gel)

explain all reactions

DC E Cw V Kpa Jsa Lua

Try first for a single antibody to

1. Gel or solid-phase most likely (probably gel)

DC E Cw V Kpa Jsa Lua

Try first for a single antibody to

1. Gel or solid-phase most likely (probably gel)

Colds: Lea, Leb, M, N, P

1. Tube testing for sure

Try first for a single antibody to

1. Tube testing for sure

Colds: Lea, Leb, M, N, P

1. Tube testing for sure

alloantibody by demonstrating lack

DC E Cw V Kpa Jsa Lua

Removing antibodies from sample by

incubation with antigen-positive RBCs

1. Gel or solid-phase most likely (probably gel)

Removing antibodies from sample by

Removal of RBC-bound antibodies

incubation with antigen-positive RBCs