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IMSE > Week 7 Lec: Antibodies > Flashcards

Week 7 Lec: Antibodies Flashcards

1
Q

True or false: The term immunoglobulin is synonymous with antibody.

A

True

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2
Q

What protein is found in the serum portion of the blood?

A

Glycoprotein

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3
Q

Percentage of polypeptides (protein) in immunoglobulins?

A

82-96%

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4
Q

Percentage of carbohydrates in immunoglobulins?

A

2-14%

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5
Q

What method is used to determine the structure of immunoglobulins?

A

serum electrophoresis

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6
Q

Materials that can be used for serum electrophoresis?

A
  1. Cellulose acetate
  2. Citrate agar
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7
Q

pH of cellulose acetate?

A

alkaline @ 8.6

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8
Q

pH of citrate agar?

A

acidic @ 6-6.3

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9
Q

Immunoglobulins appear primarily where?

A

Gamma Regions

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10
Q

Immunoglobulins are part of the ___________ branch of the immune response.

A

humoral branch

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11
Q

Immunoglobulins play an essential role during?

A

Antigen recognition

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12
Q

Immunoglobulins play an essential role in biological activities related to immune response such as?

A

opsonization and complement activation

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13
Q

Abbreviation for immunoglobulin?

A

Ig

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14
Q

Proteins in Alpha 1 Globulin?

A
  • alpha-1 antitrypsin
  • alpha-fetoprotein
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15
Q

Proteins in Alpha 2 Globulin?

A
  • ceruloplasmin
  • haptoglobin
  • alpha-2 macroglobulin
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16
Q

Proteins in Beta Globulin?

A
  • transferrin
  • hemopexin (pero hemopoietin sa notes ko? ewan)
  • complement system
  • fibrinogen
  • lipoproteins (LDL, HDL, VLDL)
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17
Q

Beta-gamma bridging is associated with what health condition?

A

liver cirrhosis

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18
Q

A toll-like spike in the gamma regions indicates what health condition?

A

multiple myeloma

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19
Q

Patients with multiple myeloma have an elevated?

A

plasma protein

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20
Q

The urine of patients with multiple myeloma contains what type of proteins?

A

Bence Jones proteins

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21
Q

Bence Jones proteins are what type of chains?

A

L chains

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22
Q

Bence Jones proteins are being secreted by?

A

malignant plasma cells

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23
Q

What happens to Bence Jones proteins heated to 60°C?

A

precipitate

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24
Q

What happens to Bence Jones proteins heated to 80°C?

A

dissolve

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25
Q

All immunoglobulin molecules are made up of a basic?

A

four-chain polypeptide unit

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26
Q

The four-chain polypeptide unit of immunoglobulins consists of?

A
  • 2 large chains, or heavy (H) chains
  • 2 smaller chains, or light (L) chains
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27
Q

The chains of immunoglobulins are held together by noncovalent forces and?

A

disulfide linkage/bond or interchain disulfide bridges

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28
Q

The structure of immunoglobulins was first described by two scientists, namely?

A

Gerald Edelman and Rodney Porter

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29
Q

Gerald Edelman and Rodney Porter won the Nobel Prize in Physiology and Medicine for describing the structure of immunoglobulins in what year?

A

1972

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30
Q

Edelman’s work centered on using the _______________ to separate out immunoglobulins on the basis of molecular weight.

A

analytical ultracentrifuge

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31
Q

Edelman’s work centered on using the analytical ultracentrifuge to separate out immunoglobulins on the basis of?

A

molecular weight

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32
Q

Edelman found that intact IgG molecules had a sedimentation coefficient of?

A

7S (Svedberg unit)

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33
Q

This indicates the sedimentation rate in an analytical ultracentrifuge.

A

Svedberg unit

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34
Q

On obtaining a purified preparation of IgG, Edelman used ______ to unfold the molecule.

A

7 M urea

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35
Q

Once unfolded, the exposed sulfhydryl bonds could be cleaved by a reducing agent, such as?

A

2-Mercapthoethanol, or simply Mercaptoethanol

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36
Q

After such treatment, the material was subjected again to ultracentrifugation, and two separate fractions were obtained. What are those fractions?

A

3.5 S and 2.2 S

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37
Q

The molecular weight of the 3.5 S fraction?

A

50,000

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38
Q

The 3.5 S fraction was designated as what chain?

A

H chain

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39
Q

The molecular weight of 2.2 S fraction?

A

22,000

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40
Q

The 2.2 S fraction was designated as what chain?

A

L chain

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41
Q

Porter’s work was based on the use of what proteolytic enzyme?

A

papain

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42
Q

Papain, which was used to cleave IgG into three pieces of about equal size, had a sedimentation coefficient of _____, and represented a molecular weight of approximately __________.

A

3.5 S; 45,000 to 50,000 d

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43
Q

Porter then subjected the immunoglobulin to?

A

Carboxymethyl cellulose ion exchange chromatography

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44
Q

After the carboxymethyl cellulose ion exchange chromatography process, the material was separated into 2 types of fragments. The one spontaneously crystallized at 4°C is called?

A

Fc fragment (for “fragment crystallizable”)

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45
Q

The remaining two identical fragments were found to have antigen-binding capacity and were named?

A

Fab fragments (fragment antigen binding)

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46
Q

How many fragments are there in papain digestion?

A

3 fragments

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47
Q

What are the 3 fragments in papain digestion?

A

2 Fab, 1 Fc

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48
Q

The Fab fragment in papain digestion consists of what chains?

A

1 light chain, 1/2 heavy chain held by disulfide bonding

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49
Q

The Fc fragment in papain digestion consists of what chains?

A

2 carboxy-terminal halves of a heavy chain

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50
Q

True or false: Fc has an antigen-binding ability.

A

False. Only Fab has an antigen-binding ability.

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51
Q

The Fc fragment is important in effector functions of immunoglobulin molecules, which include?

A

opsonization and complement fixation

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52
Q

He used pepsin to obtain additional evidence for the structure of immunoglobulins.

A

Alfred Nisonoff

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53
Q

This proteolytic enzyme was found to cleave IgG at the carboxy-terminal side of the interchain disulfide bonds.

A

pepsin

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54
Q

Pepsin was found to cleave IgG at the __________ side of the interchain disulfide bonds.

A

carboxy-terminal

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55
Q

Cleave IgG yielded?

A
  • one single fragment, molecular weight of 100,000 d, have antigen-binding ability - F(ab’)2
  • An additional fragment called FC’, which was similar to FC except that it disintegrated into several smaller pieces
56
Q

How many fragments are there in pepsin digestion?

A

2 fragments

57
Q

What are the 2 fragments in pepsin digestion?

A

1 F(ab)2, 1 Fc

58
Q

The F(ab)2 fragment in pepsin digestion consists of what chains?

A

2 light chains, 2 halves of a heavy chain

59
Q

The Fc fragment in pepsin digestion consists of what chains?

A

same with the Fc fragment in papain digestion (2 halves of a heavy chain) except that it disintegrated into several smaller pieces

60
Q

Regions where the antigen binds to the immunoglobulin?

A

Variable and Constant regions

61
Q

The region where the amino-terminal end is located?

A

Variable region

62
Q

The region where the carboxyl terminal is located?

A

Constant region

63
Q

Fab consists of what region?

A

Variable region

64
Q

Fc consists of what region?

A

Constant region

65
Q

Which immunoglobulins have 2 domains of L chain and 4 domains of H chain?

A

IgG, IgA, IgD

66
Q

What are the 2 domains of L chain and 4 domains of H chain in IgG, IgA, and IgD?

A
  • 2 domains of L chain: Variable light chain and Constant light chain
  • 4 domains of H chain: Variable heavy chain and Constant heavy chains 1, 2, and 3
67
Q

Which immunoglobulins have 2 domains of L chains and 5 domains of H chains?

A

IgE, IgM

68
Q

What are the 2 domains of L chain and 5 domains of H chain in IgE and IgM?

A
  • 2 domains of L chain: Variable light chain and Constant light chain
  • 4 domains of H chain: Variable heavy chain and Constant heavy chains 1, 2, 3, and 4
69
Q

Analysis of several Bence Jones proteins revealed that there were two main types of L chains, which were designated as?

A

kappa (κ) chains and lambda (λ) chains

70
Q

Kappa chain is coded on chromosome?

A

chromosome 2

71
Q

How many amino acids does a kappa chain have?

A

200

72
Q

Lambda chain is coded on chromosome?

A

chromosome 22

73
Q

How many amino acids does a lambda chain have?

A

220

74
Q

The region from position number 111 (the amino terminus is position number 1) is called?

A

Constant region

75
Q

The first approximately 110 amino acids at the aminoterminal end constitute which region?

A

Variable region

76
Q

True or false: The remaining amino acids after position 110 can typically be divided into three or more constant regions with very similar sequences.

A

True

77
Q

True or false: Constant regions of the H chain are unique to each class and give each immunoglobulin type its name.

A

True

78
Q

IgG has a/an?

A

Gamma chain “ɣ”

79
Q

IgM has a/an?

A

Mu chain “µ”

80
Q

IgE has a/an?

A

Epsilon chain “ɛ”

81
Q

IgD has a/an?

A

Delta chain “ẟ”

82
Q

IgA has a/an?

A

Alpha chain “α”

83
Q

A unique amino acid sequence in the H chain that is common to all immunoglobulin molecules of a given class in a given species is referred to as?

A

Isotype

84
Q

Genetic variations in the constant regions are referred to as?

A

Allotype

85
Q

Minor variations of amino acid sequences that are present in some individuals but not others are known as?

A

Allotype

86
Q

Where are allotypes seen?

A
  • In 4 IgG subclasses (IgG1, IgG2, IgG3, and IgG4)
  • In 2 IgA subclasses (IgA1 and IgA2)
  • Kappa chain of light chain
87
Q

Variations in variable regions that give individual antibody molecules specificity are referred to as?

A

Idiotype

88
Q

The segment of H chain located between the CH1 and CH2 regions is known as the?

A

Hinge Region

89
Q

The hinge region is mainly composed of an amino acid called?

A

Proline

90
Q

The high proline content of the hinge region allows for?

A

flexibility

91
Q

The ability of a hinge region to bend allows?

A

the two antigen-binding sites operate or work independently

92
Q

Hinge regions are also part or segment of?

A

heavy chains

93
Q

Immunoglobulins that have a hinge region?

A

IgG, IgA, IgD

94
Q

Immunoglobulins that DO NOT have a hinge region?

A

IgE, IgM

95
Q

The ______________ will determine the number of antigen-binding sites.

A

valence of immunoglobulin

96
Q

True or false: The more valence in antibodies, the more antigen-binding sites there are.

A

True

97
Q

3 basic immunoglobulin structures?

A
  • Monomer
  • Dimer
  • Polymer
98
Q

Immunoglobulin structure that has 2 antigen binding sites?

A

Monomer

99
Q

Immunoglobulin/s having a monomer structure?

A
  • IgG
  • IgA1/serum IgA
  • IgD
  • IgE
  • IgM (specifically only on the surface of B cells)
100
Q

Immunoglobulin structure that has 4 antigen binding sites?

A

Dimer

101
Q

Immunoglobulin/s having a dimer structure?

A

IgA2/secetory IgA

102
Q

Immunoglobulin structure that has more than 4 antigen binding sites?

A

Polymer or Pentameric

103
Q

Immunoglobulin/s having a polymer structure?

A

IgM because it has a valence of 10

104
Q

True or false: Attempts to explain the specificity of antibody for a particular antigen began long before the actual structure of immunoglobulins was discovered.

A

True

105
Q

The 4 theories of antibody diversity?

A
  • Ehrlich Side-Chain Theory
  • Clonal Selection Theory
  • Template Theory
  • Selective Theory
106
Q

The theory included as part of Erlich Side-Chain Theory?

A

Selective Theory

107
Q

Theory involved in the lock and key model of antigen and antibody.

A

Erlich Side-Chain Theory

108
Q

He postulated that certain cells had specific surface receptors for antigen that were present before contact with antigen occurred.

A

Ehrlich

109
Q

Determine what theory is being described: Once antigen was introduced, it would select the cell with the proper receptors, combination would take place, and then receptors would break off and enter the circulation as antibody molecules.

A

Erlich Side-Chain Theory

110
Q

In the 1950s, they independently supported the idea of a clonal selection process for antibody formation.

A

Niels Jerne and Macfarlane Burnet

111
Q

When did Niels Jerne and Macfarlane Burnet independently supported the idea of a clonal selection process for antibody formation?

A

1950s

112
Q

Most acceptable theory in antibody diversity.

A

Clonal Selection Theory

113
Q

Determine what theory is being described: The key premise is that individual lymphocytes are genetically preprogrammed to produce one type of immunoglobulin and that a specific antigen finds or selects those particular cells capable of responding to it, causing them to proliferate.

A

Clonal Selection Theory

114
Q

Who created the Template Theory?

A

Felix Haurowitz

115
Q

Second major theory of antibody diversity in 1930’s?

A

Template Theory

116
Q

Determine what theory is being described: Antibody-producing cells are capable of synthesizing a generalized type of antibody, and when contact with an antigen occurs, the antigen serves as a mold or template and alters protein synthesis so the antibody with a specific fit is made. This specific antibody produced enters the circulation, while antigen remains behind to direct further synthesis.

A

Template Theory

117
Q

They discovered a technique to produce antibody arising from a single B cell.

A

George Kohler and Cesar Milstein

118
Q

George Kohler and Cesar Milstein discovered a technique to produce antibody arising from a single B cell, which is called?

A

Monoclonal Antibody

119
Q

In result, the discovery of monoclonal antibody revolutionized?

A

serological testing

120
Q

Kohler and Milstein’s technique fuses an __________ with a __________ that can be grown indefinitely in the laboratory.

A

activated B cell, myeloma cell

121
Q

Myeloma cells lacks an enzyme called?

A

HGPRT or Hypoxanthine-guanine phosphoribosyltransferase

122
Q

It is an important enzyme to synthesize nucleotides from hypoxanthine and thymidine.

A

HGPRT or Hypoxanthine-guanine phosphoribosyltransferase

123
Q

They are cancerous plasma cells.

A

Myeloma cells

124
Q

Method used to produce monoclonal antibodies?

A

Hybridoma Production or Hybridoma Technique

125
Q

It is a high-end type of antibody.

A

Monoclonal Antibody

126
Q

True or false: The knowledge that B cells are genetically preprogrammed to synthesize very specific antibody has been used in developing monoclonal antibodies for diagnostic testing.

A

True

127
Q

Hybridoma production: One pathway, which builds DNA from degradation of old nucleic acids, is?

A

blocked

128
Q

Hybridoma production: The other pathway, which makes DNA from new nucleotides, is blocked by the presence of?

A

aminopterin

129
Q

Hybridoma production: The remaining hybridoma cells are diluted out and placed in ___________, where they are allowed to grow.

A

microtiter wells

130
Q

Hybridoma production: Each well, containing one clone, is then screened for the presence of the desired antibody by?

A

removing the supernatant

131
Q

Hybridoma production: Once identified, a hybridoma is capable of being maintained in cell culture indefinitely, and it produces a permanent and uniform supply of monoclonal antibody that reacts with a?

A

single epitope

132
Q

Fusing medium for plasma cells and myeloma cells?

A

PEG - Polyethylene glycol

133
Q

True or false: During Hybridoma production, a large percentage of cells will fuse.

A

False. Only a small percentage of cells will actually fuse.

134
Q

In Hybridoma production, the fused cells are placed in a selective culture media called?

A

HAT Medium (hypoxanthine-aminopterin-thymidine medium)

135
Q

Why will myeloma cells die after placing the fused cells in HAT medium?

A

Because myeloma cells lack the enzyme HGPRT, they cannot utilize hypoxanthine and thymidine

136
Q

Why will spleen cells die after placing the fused cells in HAT medium?

A

Because normal B cells cannot be maintained continuously in cell culture

137
Q

Which are the only cells that will grow after Hybridoma production?

A

Hybridoma cells

IMSE (9 decks)

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