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BB Lec > ANTIGLOBULIN TESTING > Flashcards

ANTIGLOBULIN TESTING Flashcards

1
Q

It obtained from immunized nonhuman species bind to human globulins such as IgG or complement either free in serum or attached to antigens on red blood cells (RBC’s).

A

ANTIHUMAN GLOBULIN TEST

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2
Q

It detect IgG or complement-sensitized RBCs?

A

ANTIHUMAN GLOBULIN TEST

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3
Q

What is the 2 MAJOR TYPES OF BLOOD GROUP ANTIBODIES

A

IgM, IgG

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4
Q

Bind to corresponding antigen and directly agglutinate RBCs suspended in saline

A

IgM

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5
Q

It is a pentamer, and is considered to be the best agglutinin

A

IgM

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6
Q

“nonagglutinating” or “incomplete antibodies” – because their single monomer structure is too small to directly agglutinate sensitized RBCs

A

IgG

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7
Q

precipitin so it produces small clumps as compared to agglutination.

A

IgG

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8
Q

1945, they described the use of the antiglobulin test for the detection of weak and nonagglutinating Rh antibodies in serum.

A

Coombs, Mourant, and Race

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9
Q

1946 ,Coombs and coworkers described the use of ?

A

AHG to detect in vivo sensitization of the RBCs of babies suffering from HDFN.

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10
Q

In 1908, Moreschi described the principle of the

A

Anti-globulin test

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11
Q

In 1908 - Moreschi described the principle of the Anti-globulin test which included what blood group antibodies and associated antigen.

A

Kell blood group antibodies and associated antigen

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12
Q

Injection of human serum into rabbits to produce antihuman serum

A

Coombs procedure

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13
Q

In Coombs procedure the one we see in this procedure is?

A

Hemagglutination

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14
Q

Result of RBCs has been sensitized by an antibody

A

Hemagglutination

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15
Q

In 1947 - Coombs and Mourant demonstrated that the antibody activity that detected

A

Rh

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16
Q

Coombs and Mourant demonstrated that the antibody activity that detected Rh
o antibodies were associated with the ?

A

anti-gamma globulin fraction

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17
Q

In 1951 (?) Observed that different reaction patterns were obtained when dilutions of AHG were used to test cells sensitized with warm as compared to cold antibodies.

A

Dacie

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18
Q

Warm reacting antibodies

A

IgG

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19
Q

Cold reacting antibodies

A

IgM

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20
Q

The Antiglobulin test can be used to detect RBCs sensitized with?

A

IgG alloantibodies, IgG autoantibodies, and complement components.

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21
Q

Sensitization can occur either in?

A

Vivo and vitro

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22
Q

Ag-Ab binding happened inside the body? In vivo or in vitro?

A

vivo

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23
Q

outside the body? In vivo or in vitro?

A

vitro

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24
Q

is detected by a one stage procedure because their already sensitized RBC in body.

A

vivo sensitization

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25
Q

vivo sensitization is detected by?

A

Direct antiglobulin test (DAT)

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26
Q

The use of AHG to detect in (?) sensitization of RBCs is a two-stage technique.

A

vitro sensitization

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27
Q

vitro sensitization is detected by

A

Indirect antiglobulin test (IAT)

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28
Q

What is the 2 procedures in vitro sensitization?

A

The 2 procedures:
- We need to incubate them to 37 degrees Celsius because they are IgG.
- Add you anti-human globulin reagent to demonstrated if they can agglutinate.

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29
Q

Defined by FDA and CBER (Center for Biologics Evaluation and Research)

A

ANTIHUMAN GLOBULIN REAGENTS

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30
Q

FDA and CBER transcribe?

A

Food and Drug Administration, Center for Biologics Evaluation and Research

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31
Q

ANTIHUMAN GLOBULIN REAGENTS 2 types?

A

POLYSPECIFIC, MONOSPECIFIC

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32
Q

is the one we check because it presents in all pathway.

A
  • C3
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33
Q

Antibody to human IgG and to the C3d component of the human complement, contains more than 1 type of antibody.

A

POLYSPECIFIC

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34
Q

POLYSPECIFIC commonly used?

A

Anti-C3b, anti- C4b and anti-C4d may also be present

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35
Q
  • contain only one antibody specificity: either anti IgG or antibody to specific complement components (C3b or C3d)
A

MONOSPECIFIC

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36
Q

Monospecific commonly used?

A

: anti-IgG and anti-C3b, anti-C3d

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37
Q

o No anticomplement activity
o Contain antibodies specific for the Fc fragment of the gamma heavy chain of the IgG molecule.
o IgG will bind to Fc portion

A

Anti-IgG

38
Q

Anticomplement reagents?

A

Anti-C3b and anti-C3d reagents

39
Q

Often a blend of monoclonal anti-C3b and monoclonal C3d

A
  • Anticomplement
40
Q

Complement is the start of the reaction that is common in 3 pathways

A

classical, alternative, mannose)

41
Q

foreign antigen to the rabbits

A
  • Human globulin
42
Q
  • Many Ag to bind
A

POLYCLONAL ANTIBODIES

43
Q
  • Mixture of antibodies from different plasma cell clones.
  • Directed against multiple types of epitopes.
A

POLYCLONAL ANTIBODIES

44
Q

determinant site that is found in the antigen to be able to recognize by the antibody

A

o Epitopes

45
Q
  • 1 Ag can only be bind.
  • Derived from one clone of plasma cells and recognize a single epitope.
A

MONOCLONAL ANTIBODIES

46
Q

can be used to produce monoclonal antiglobulin serum

A
  • Hybridoma technology
47
Q

POLYCLONAL AHG PRODUCTION usually prepared using?

A

rabbits

48
Q

IN POLYCLONAL AHG PRODUCTION, Large volumes of antibody are required (?) and (?) may be used

A

Sheep or Goats

49
Q
  • If the rabbit is injected with a complement, it will produce
A

Monospecific Polyclonal Anti-C3b, Anti-C3d.

50
Q
  • If the rabbit is injected with an IgG, it will produce
A

Monospecific Polyclonal Anti-IgG.

51
Q

When we mixed both Monospecific Polyclonal Anti-C3b, Anti-C3d and Monospecific Polyclonal Anti-IgG together, it can produce

A

Antihuman Globulin Polyspecific Polyclonal Blend

52
Q

means there is only 1 Ab but can detect many epitopes (reacts)

A

Monospecific Polyclonal

53
Q
  • Recognize multiple epitopes on any one antigen.
A

POLYCLONAL ANTIBODIES

54
Q

contains a heterogenous complex mixture of antibodies of different affinity.

A
  • Serum obtained
55
Q

made up mainly of IgG subclass. (4 subclasses of IgG)

A
  • Polyclonals
56
Q
  • often used to generate polyclonal antibodies that target unique epitopes, especially for protein families of high homology
A
  • Peptide immunogens
57
Q

MONOCLONAL AHG PRODUCTION also called?

A

hybridoma technology

58
Q

MONOCLONAL AHG PRODUCTION was discovered by?

A

Kohler and Milstein

59
Q

in monoclona AHG production, Immunization of laboratory animals usually use is (?) with purified human globulin.

A

mice

60
Q

For HYBRIDOMA to form we need to incubate it to

A

HAT medium (hypoxanthine–aminopterin–thymidine)

61
Q

IN PREPARING ANTI COMPLEMENT in MONOCLONAL AHG PRODUCTION we will have?

A

Monoclonal anti-C3b, d, or g (1 only Ab target)

62
Q

IN PREPARING ANTI-IGG in MONOCLONAL AHG PRODUCTION we will have?

A

Monoclonal ant-IgG (1 only Ab target)

63
Q

If we mix the anti-C3b,d,g and anti-IgG, we will have an

A

anti-human globulin (AHG) polyspecific monoclonal blend

64
Q
  • Contains only one antibody specificity
A

MONOSPECIFIC AHG

65
Q

is produced as a monoclonal, polyclonal or blended formula.

A
  • Monospecific anti-IgG
66
Q
  • Anti-IgG: Majority are mixture of (?) and (?) subclasses
A

IgG1 and IgG3

67
Q
  • High specificity
  • Detects only one epitope on the antigen.
  • Consist of only one antibody subtype (e.g. IgG1, IgG2, IgG3, iGG4)
A

MONOCLONAL ANTIBODIES

68
Q
  • Antibody molecules and complement components are
A

globulins

69
Q

reacts with human globulin molecules, either bound to RBCs or free in serum. Washed RBCs coated with human globulin are agglutinated.

A

AHG

70
Q

Detects in vivo sensitization of RBCs with IgG or complement components. Clinical conditions that can result in in vivo coating of RBCs with antibody or complement

A
  • DAT
71
Q

Clinical conditions that can result in in vivo coating of RBCs with antibody or complement are:

A

o HDN
o HTR
o Autoimmune hemolytic anemia and drug-induced hemolytic anemia

72
Q

RBC have metabolites of the drugs attached which usually cause by ?

A

ibuprofen

73
Q

o One drop of 3% to 5% suspension of washed RBCs with polyspecific (anti-IgG, anti-C3d) reagents.

A
  • DAT Panel
74
Q

Anti-IgG (+) and Anti-C3d (+)

A

67% of Warm Autoimmune Hemolytic Anemia

75
Q

Anti-IgG (+) and Anti-C3d (-)

A

20% Warm Autoimmune Hemolytic Anemia cases

76
Q

Anti-IgG (-) and Anti-C3d (+)

A

(13%) of Cold Hemagglutinin disease (CHD), Paroxysmal Cold Hemoglobinuria (PCH), Warm Autoimmune Hemolytic Anemia (WAIHA)

77
Q

In vivo sensitization: Maternal Ab coting fetal RBC

A

HDN

78
Q

In vivo sensitization: Recipient Ab coating donor RBC

A

HTR

79
Q

In vivo sensitization: Autoantibody coting individual RBC

A

AIHA

80
Q

In Hemolytic Disease of the Newborn (HDN), what we check for DAT will be

A

Fetal RBC

81
Q

In Hemolytic Transfusion Reaction (HTR) we will be checking the

A

Recipient blood

82
Q
  • Own antibodies are attacking its own cell
A

Autoantibody

83
Q
  • performed to determine in vitro sensitization of RBCs and is used in the following situations
A

INDIRECT ANTIGLOBULIN TEST (IAT)

84
Q

o Detection of incomplete (non-agglutinating) antibodies to potential donor RBCs (compatibility testing) or to screening cells (antibody screen) in serum

A

INDIRECT ANTIGLOBULIN TEST (IAT)

85
Q

Ab detection

A

copatibility testing, Ab screening

86
Q

Ab identification

A

Ab panel

87
Q

Ab titrataion

A

Rh Ab titer

88
Q

RBC phenotype

A

RBC Ag detection

89
Q
  • Cells suspended in saline:
A

INCUBATION TIME

90
Q

incubation times vary between (?) if there is no reaction medium

A

30-120 minutes

91
Q

shortened the Incubation time to 10 to 15 minutes

A

Low-Ionic-Strength Solution, Polyethylene Glycol

BB Lec (4 decks)

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