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Introduction to Antibody Identification Justin R. Rhees, M.S., MLS(ASCP) CM ,SBB CM University of Utah Medical Laboratory Sciences
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Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Jun 13, 2019

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Page 1: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Introduction to

Antibody

Identification

Justin R. Rhees, M.S., MLS(ASCP)CM,SBBCM

University of Utah

Medical Laboratory Sciences

Page 2: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Objectives

After this presentation the learner will be able to correctly:

1. State the clinical utility of antibody identification.

2. Describe the principle and procedure of the antibody identification tests.

3. Explain heterozygosity and homozygosity as they apply to antibody identification.

4. List allelic pairs in the following blood group systems: Rh, Duffy, Kidd, MNSs.

5. Given patient test results, work through the antibody identification process.

Page 3: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Screening and Identification

• Antibody screening is required: 1. Any intended recipient of transfused blood.

2. Prenatal testing for obstetric patients: evaluates risk of HDFN and candidacy for RhIg

3. Donors of allogeneic blood and blood products and stem/progenitor cells.

• Further testing (Antibody ID) is required: 1. If the screen is newly positive

2. New alloantibody(ies) suspected

Page 4: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification

Page 5: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Methods for ABID

• Antibody identification (ABID) can be performed in the following media:

– Traditional tube method

– Gel Technique

– Solid Phase Technique

Page 6: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody exclusion

• An exclusion procedure can be undertaken through observation of antigens present on reagent cells with which patient sera did not react.

• This means the patient’s antibodies are not likely directed against the antigens present on those cells.

Page 7: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Procedure

• Go to the first panel cell with a negative reaction, “rule out” or exclude the specificities of antibodies directed against antigens present on the cell.

– (Rule out when the antigen is positive and the patient did not react)

– Some antibodies demonstrate dosage.

Page 8: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

• Some antibodies may react so weakly with antigens with heterozygous expression, they might not be detected.

• For antibodies in the following blood groups, it may be prudent to rule out with panel cells that have a homozygous expression of antigen: – Rhesus (excluding D)

– Kidd

– Duffy

– MNSs

Page 9: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Page 10: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 11: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 12: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 13: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 14: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 15: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 16: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 17: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Page 18: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 19: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 20: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 21: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 22: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 23: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 24: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

Mother Father

Page 25: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Dosage

• Anti-Jka may not react with a heterozygous “single dose” cell

• It may only react with a cell that has “double the dose” of Jka antigens

Page 26: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Allelic pairs Rh System C, c

E, e

Duffy System Fya, Fyb

Kidd System Jka, Jkb

MNSs System M, N

S, s

Page 27: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 +

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 +

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + +

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + +

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 +

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 +

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0

8 + + + + + 0 + + + + + 0 + + 0 +

AC

Page 28: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 29: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 30: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 31: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 32: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 33: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 34: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 35: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 36: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 37: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 38: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 39: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 40: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 41: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 42: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 43: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 44: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 45: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 46: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 47: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

The next step

• What alloantibody or alloantibodies have not been ruled out?

Anti-E

Anti-Fya

Which of the following is or are most likely?

Look closely at the pattern of reactivity.

Page 48: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 49: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 50: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 51: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Confirmation steps

• Anti-E is the most likely antibody reacting

• However, we still have not ruled out anti-Fya

• The patient could have anti-Fya underlying the reactions of anti-E

We need to select another cell that is

E antigen negative, and Fy(a+b-)

HOMOZYGOUS for Duffy A

Page 52: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Selected Cell from a different panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + 0 + 0 + 0 + 0 + + + + 0 +

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 +

3 + 0 + 0 + + + + 0 + 0 + 0 + + +

4 + + + + + 0 + 0 0 + + 0 + 0 + +

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 +

6 0 0 + + + 0 + 0 0 + 0 0 + 0 0 +

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0

8 0 + + 0 + 0 + + + + + 0 + + 0 +

AC

Page 53: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Selected Cell from a different panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + 0 + 0 + 0 + 0 + + + + 0 +

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 +

3 + 0 + 0 + + + + 0 + 0 + 0 + + +

4 + + + + + 0 + 0 0 + + 0 + 0 + +

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 +

6 0 0 + + + 0 + 0 0 + 0 0 + 0 0 +

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0

8 0 + + 0 + 0 + + + + + 0 + + 0 +

AC

Page 54: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Selected Cell from a different panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + 0 + 0 + 0 + 0 + + + + 0 +

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 +

3 + 0 + 0 + + + + 0 + 0 + 0 + + + 0 0 ✓

4 + + + + + 0 + 0 0 + + 0 + 0 + +

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 +

6 0 0 + + + 0 + 0 0 + 0 0 + 0 0 +

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0

8 0 + + 0 + 0 + + + + + 0 + + 0 +

AC

Page 55: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Selected Cell from a different panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + 0 + 0 + 0 + 0 + + + + 0 +

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 +

3 + 0 + 0 + + + + 0 + 0 + 0 + + + 0 0 ✓

4 + + + + + 0 + 0 0 + + 0 + 0 + +

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 +

6 0 0 + + + 0 + 0 0 + 0 0 + 0 0 +

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0

8 0 + + 0 + 0 + + + + + 0 + + 0 +

AC

Page 56: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Rule of 3

• Criteria:

– At least 3 panel cells with E antigen reacted (positive result) with patient’s sample

– At least 3 panel cells lacking E antigen did not react (negative result) with the patient’s sample

• Does our example fulfill these criteria?

Page 57: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 58: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification Panel Rh-Hr Kell Duffy Kidd P MNSs Results

D C c E e K k Fya Fyb

Jka Jkb P1 M N S s 37 AHG CC

1 + 0 + + + 0 + 0 0 + + + + + 0 + 0 2+

2 + 0 + 0 + 0 + 0 0 + 0 + + 0 0 + 0 0 ✓

3 + 0 + + 0 0 + 0 0 + 0 + 0 + + + 0 2+

4 0 + 0 + + 0 + 0 0 + + 0 + 0 + + 0 0 ✓

5 0 0 + + + 0 + 0 0 0 + + 0 + 0 + 0 2+

6 0 0 + 0 + 0 + 0 0 + 0 0 + 0 0 + 0 0 ✓

7 0 0 + 0 + + + 0 + 0 + 0 0 + + 0 0 0 ✓

8 + + + + + 0 + + + + + 0 + + 0 + 0 2+

AC

Page 59: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Yes!

• Positive for E, patient reacted:

Cells 1, 3, 5, 8

• Negative for E, patient did not react:

Cells 2, 6, 7

Cell 4 can’t be used for this because it is E antigen positive

Page 60: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Rule of 3

• At least 3 true positives and 3 true negatives:

Following this rule gives us a P value of 0.05

95% chance that the antibody we have identified is correct.

Page 61: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Result

• Anti-E identified. All other clinically significant alloantibodies have been ruled out.

• Donor units lacking E antigen should appear crossmatch compatible through the indirect antiglobulin test (IAT).

Page 62: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

In conclusion

• Rules for determining how antibody identification is performed are determined by the facility.

Rule of 2?

2 in 3 out?

Heterozygous ok if 2

Etc.

Page 63: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

In conclusion

• Some patient antibodies may be so weak, they are only detected with a homozygous cell.

• Antibodies may behave differently in different media: – Solid phase?

– Gel?

– Tube technique: • PeG?

• LISS?

Page 64: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

In conclusion

• With multiple alloantibodies, it is important to prove them independently of each other.

• The process of antibody identification can take several hours in order to determine the specificity of the alloantibody(ies).

Page 65: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification

Example 1:

Cold Reactive Alloantibody

Page 66: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification

Example 2:

Multiple Warm Reactive

Alloantibodies

Page 67: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Antibody Identification

Example 3:

Alloantibody demonstrating

dosage

Page 68: Introduction to Antibody Identification - ARUP Scientific ...arup.utah.edu/media/AntibodyIdentification/Introduction to Antibody Identification.pdf · Introduction to Antibody Identification

Thank you!


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