- monoclonal IgM antibody reactive at room temp - monoclonal polyclonal blend includes IgG antibody for weak D detection

Antibody Identification Flashcards by Savannah Whitmire

agency that controls blood bank reagents

Bureau of Biologics of FDA

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types of blood bank reagents

high protein, monoclonal, chemically modified, human-derived

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ABO reagents

commercially prepared anti-A, anti-B, anti-A,B, anti-A1, A1 and B cells

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RH reagents

monoclonal IgM antibody reactive at room temp

- monoclonal polyclonal blend includes IgG antibody for weak D detection

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uses of indirect antiglobulin test

antibody screen/panel
crossmatch
weak D testing
leukocyte and platelet antibody test
detection of autoantibodies

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uses of direct antiglobulin test

detection of autoantibodies
HDFN
transfusion reactions
RBC sensitization by drugs

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examples of enhancement media

LISS
PEG
22% bovine serum albumin
gel column agglutination
solid-phase technology

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what should be suspected if autocontrol is positive but DAT is negative?

antibodies to LISS and PEG enhancement media ingredients

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mechanism: LISS

low ionic strength saline; reduced ionic strength in comparison to normal saline

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which enhancement media(s) is used to enhance reactivity and decrease incubation time for antibody identification?

LISS, PeG

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which enhancement media may enhance autoantibodies?

LISS

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mechanism: PeG

polyethylene glycol; linear polymer that is water-soluble; steric exclusion of water molecules in the diluent

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what enhancement may enhance warm autoantibodies?

PEG

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what enhancement enhances detection of clinically significant antibodies along with decreasing detection of clinically insignificant antibodies?

PEG

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mechanism: 22% bovine serum albumin

helps reduce repulsive charges between red cells during antibody detection, promoting agglutination

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what enhancement may enhance cold autoantibodies?

22% bovine serum albumin

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principle use of enhancement media

reduce incubation time and increase reaction strength

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what media enhances linked lattice formation by reducing net negative charge of red cells, allowing the cells to come closer together?

albumin

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what media enhances antibody uptake by reducing zeta potential and allowing increased attraction between positively charged antibodies and negatively charged red cells?

LISS

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what media enhances antibody-antigen binding by excluding water from around the red cells, effectively concentrating the antibody and favoring binding to its target antigen?

PEG

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what media enhances linked lattice formation by decreasing the net negative charge of the red cells by removing sialic acid residues to allow the red cells to come closer together?

enzyme treatment

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what media enhances formation of the linked lattice by neutralizing the negative charges of the sialic acid residues on the red cells, allowing red cells to get closer together?

polybrene

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media that can enhance reactivity of weak Kidd and Duffy antibodies

PEG

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enhancement that may not pick up IgM antibodies such as ABO or Lewis

PEG

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enhancement that can result in spontaneous agglutination

polybrene

avidity

speed at which standardized endpoint is attained (strength)

titer

reciprocal of greatest dilution that gives 1+ agglutination (amount)

requirements at reagent arrival

dated and stored at optimal temp | expiration date and appearance noted and recorded

QC of antigen typing sera

heterozygous positive control | negative control

naturally occurring antibody common class

IgM

immune stimulated antibody common class

IgG

antibodies to substances in the environment that are antigenically similar to red cell antigens are ___ (naturally occuring or immune stimulated)

naturally occurring

definition: clinically significant antibody

causes HDFN or HTR or noticeable decrease in transfused red cell survival

preanalytical patient medical history considerations in antibody identification

transfusion, pregnancy, disease, drugs, rare phenotype

analytical considerations in antibody identification

specimen requirements, reagents, serological procedures, factors affecting antibody identification

p value requirement of antibody identification

<0.05

causes of positive autocontrol

autoantibodies (Warm and cold) recent transfusion/DHTR drug-induced issues passively acquired antibodies

source of ficin

figs

source of papain

papaya

source of trypsin

pig stomach

source of bromelin

pineapple

antigens destroyed by enzyme treatment

M, N, Fya, Fyb, Xga, JMH, Ch, Rg

antigens enhanced by enzyme treatment

Rh, Lewis, Kidd, P, I

mechanism of enzyme treatment

cleave membrane glycoproteins and sialic acid residues that link amino acid chains together

antigens unaffected by enzyme treatment

Kell Diego Colton

mechanism of thiol reagents

disrupt disulfide bonds that connect J chain of IgM molecule; denatures IgM molecules so they can't agglutinate or bind complement

thiol reagents

DTT 2-ME AET

reagent used to distinguish IgM from IgG

thiol

what enzyme denatures CD38 antigen

thiol

antibody seen in Dara patients

anti-CD38

thiol reagent effect on Kell antigens

destroys

mechanism of ZZAP

denatures DTT sensitive antigens along with antigens that are sensitive to enzymes

mechanism of chloroquine diphosphate

dissociate IgG from red cell membrane; weakens class I HLA antigen expression along with other antigens including Rh

reagent used to remove bound antibody so cells can be antigen typed using antisera that require IAT testing

glycine acid/EDTA, chloroquine diphosphate

what antibodies can be enhanced by lowering pH to 6.5

anti-M, some anti-U

how to lower pH to 6.5

1 volume 0.1N HCl to 9 volumes serum

lowering pH to <6.0 can decrease or diminish reactivity of what blood groups

Rh, Duffy, Kidd

Lewis inhibition substance

saliva of individual with Lewis gene

P1 inhibition substance

hydatid cyst fluid, pigeon eggs

Sda inhibition substance

urine from Sda positive individual

Chido and Rodgers inhibition substance

plasma from Ch+Rh+ individual

what is required to interpret results from inhibition techniques?

overall test result is nonreactive and dilution control is negative

mechanism: adsorption

antibodies removed from patient serum by adsorbing onto red cells which express the corresponding antigen

normal serum:cell ratio for adsorption

1:1

mechanism: elution

bound antibody is released by changing of thermodynamics to recover bound antibody into usable form

heat/freeze-thaw elution use

ABO incompatibility from ABO HDFN

acid/organic solvent elution use

warm autoantibodies and alloantibodies

mechanism: titration

testing twofold serial dilutions of patient's serum with preselected red blood cells to provide relatively how much antibody is present in patient's serum or given strength of red blood cell antigen expression