RELATIONSHIP BETWEEN IMMUNOLOGY AND IMMUNOHEMATOLOGY Part 2

__________________ - Enzyme that cuts the monomer exactly at the Hinge region, monomer is cut into 3 parts: 2 FAB and 1 Fc region.

__________________ - Enzyme that cuts the monomer into 2 parts

 Papain - Cut monomer exactly at the Hinge region, monomer is cut into 3 parts: 2 FAB and 1 Fc region.

 Pepsin - Cut monomer into 2 parts

Factors that influence Antigen-Antibody Reaction (5)

Antigen-Antibody Ratio

pH

Temperature

Immunoglobulin type

Centrifugation

In the Ag-Ab ration. the dosage is affected by:

__________________ – stronger AbAg reaction

__________________ – weaker AbAg reaction

- Homozygous concentration – stronger AbAg reaction

- Heterozygous concentration – weaker AbAg reaction

pH where Anti-M and Anti-PR reacts best at _____________

Acidic pH

Enhance agglutination reactions because it decreases reaction time by increasing the gravitational forces of the reactants.

CENTRIFUGATION

Temperature

To detect IgG –

 To detect IgM –

To detect IgG – 37C

To detect IgM – room temperature

force repulsion between red cells due to their difference in electrostatic charges surrounding the cations

Zeta Potential

Reagents that enhance the detection of IgG antibodies by increasing their reactivity.

Reduces the zeta potential

ENHANCEMENT MEDIUM / POTENTIATOR

Increases the dielectric constant (a measure of electrical conductivity), which then reduces the zeta potential of the RBC.

Protein Media

Most commonly used Protein Media

22 or 30% bovine serum albumin

Protein media that removes the water molecule in the environment

More effective than albumin, LISS for detection of weak antibodies

Polyethylene glycol (PEG)

Most commonly used Potentiator

Decreases the ionic strength of a reaction medium, which reduces the zeta potential and therefore allows antibodies to react more efficiently with RBC membrane antigens

Most commonly used enhancement medium of IAT due to its short test incubation time

Low ionic strength sollution

Potentiator that:

 Target the sialic component, protein molecule

 Enhances: Rh, Kidd, P, Lewis, and I antigens

 Destroys: Fya, Fyb, M, N, S antigens

Proteolytic Enzymes

Potentiator that Detects antibodies and complement proteins that have attached to the RBC

 Involved the injection of human serum into rabbits to produce antihuman serum

Antihuman Globulin Test

Antiglobulin Test that detects in vivo sensitization of RBC with IgG/complement components

Direct Antihuman Globulin (DAT)

Clinical Conditions for DAT

HDFN

HTR

AIHA

Hemolytic Disease of the Fetus and Newborn

Hemolytic Transfusion Reaction

Autoimmune Hemolytic Anemia

Sample of Choice for DAT Test

EDTA Blood

DAT Cell that is used to detect and validate the negative results. Also to check if you really add the AHG reagent

Check Cells

Detects in vitro sensitization of RBC using specimen serum

Indirect Antihuman Globulin

IAT is used for the following (3)

AD

AI

AT\

RCP

CM

Antibody Detection

Antibody Identification

Antibody Titration

Rec Cell Phenotyping

Crossmatching

AHG Reagents (2)

Polyspecific AHG

Monospecific AHG

______________________ - AHG Reagent that contains antibody to human IgG and to the C3d component of human complement.

______________________ - AHG Reagent that contains only one antibody specificity; either anti-IgG or antibody to specific components of complement, such C3b or C3d.

 Polyspecific AHG - Contain antibody to human IgG and to the C3d component of human complement.

 Monospecific AHG - Contain only one antibody specificity; either anti-IgG or antibody to specific components of complement, such C3b or C3d.

AHG Prep where the Mixture of antibodies from different plasma cell clones. - Recognize different antigenic determinants (epitopes), or the same portion of the antigen but with different affinities.

Polyclonal

AHG Prep Derived from one clone of plasma cells and recognize a single epitope. - Mice

Monoclonal

POSITIVE REACTIONS IN BLOOD BANK (AHP)

Agglutination

Hemolysis

Precipitation

Antibody-mediated clumping of particles that express antigen on the surface.

Agglutination

Rupture of red cells with release of intracellular hemoglobin - Complement fixation happens

Hemolysis

Formation of an insoluble complex when soluble antigen reacts with soluble antibody.

Precipitation

disappears with addition or normal saline solution (positive result in blood bank)

Rouleaux

 Small agglutinated within predominantly agglutinated red cells

Mixed Field