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Molecular > Molecular Methods Dr. Ford > Flashcards

Molecular Methods Dr. Ford Flashcards

1
Q

Purpose of auxotophy

A

Nutrient MUST be included in medium.

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2
Q

4 principles for cell culture

A

Food, shelter, climate control, and companionship

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3
Q

What does having an antibiotic do for cell culture?

A

Drug resistance; only cells you will want will grow in presence of antibiotic. Creates SELECTION

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4
Q

What is a feeder cell layer?

A

Layer of fibroblast

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5
Q

What are factors of climate control?

A

Humidity, pH (CO2 is a regulator; acts as a buffer), oxygen (anaerobic vs aerobic, and light

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6
Q

What is a symbiotic relationship?

A

Some cells need other cells to survive; one type of a cell may make nutrients the other cell needs

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7
Q

What is contact inhibition?

A

Change in characteristics due to touching

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8
Q

What is quorum sensing?

A

Cells figuring out if a neighboring cell is friend or foe

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9
Q

Some cells can dilute into a single cell and will form colony ie

A

Colony formation

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10
Q

What are the cells for cell mediated immunity?

A

T cells

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11
Q

Humoral immunity is what type of cells?

A

B cells; makes antibodies but not as much as plasma cells

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12
Q

Plasma cells useful for:

A

Differentiated B cells with pronounced ER to make lots of antibodies

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13
Q

T cells, B cells, and plasma cells all work to:

A

Identify other types of cells

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14
Q

Characteristics of an antibody:

A

“Y” shape= 4 proteins; 2 heavy long chains and 2 light shorter chains. Held together with 4 DISULFIDE bonds

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15
Q

Disulfide bonds between cysteines require what type of environment?

A

OXIDIZING; ie to create antibodies

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16
Q

What are some examples of oxidizing environments?

A

Periplasmic space
OM in prokaryotes
ER lumen
Peroxisomes in eukaryotes

17
Q

What does a reducing environment do to an antibody?

A

Creates a non-stable antibody ie destroys

18
Q

What is a conserved property for an antibody?

A

Conserved immunoglobulin domain and conserved Beta strands

19
Q

What is variable in an antibody?

A

3 binding loops. These loops bind antigens and different from antibody to antibody

20
Q

What kind of bonds for antibody/antigen binding and what properties as well?

A

Non-covalent (attractive but not permanent) and size and shape are complimentary for binding

21
Q

What are epitopes?

A

The “fingers” that interact with the antibody. Need to be aware of what epitope and the state of the protein ie folded vs non-folded

22
Q

Mono = 1 epitope only. Expensive of non-expensive?

A

Expensive

23
Q

Poly = many different antibodies that bind to different epitopes. Expensive or cheap?

A

Cheap

24
Q

What are MAbs and what are they used for?

A

Monoclonal antibodies. Herceptin is a MAb shown to bind to HER2 receptors to prevent growth. Targets different aspects of the immune system and prevent inflammation

25
Q

What is the ‘IP’ in ChIP

A 
Immuno = antibodies
P = precipitation = heavy agarose beads (sink) or magnetic for stickiness; will remove out the of system
26
Q

What is ‘Ch’ in ChIP

A

Chromosome

27
Q

How does ChIP function?

A

Takes cells and make a covalent interaction with DNA and proteins. Antibody is specific for protein of interest and follows and captures all DNA cross-linked to the antigenic protein.

28
Q

What does formaldehyde do in ChIP?

A

Formaldehyde connects DNA and protein at non-covalent interactions and also releases covalent binds to see what was captured (DNA)

29
Q

Some examples ChIP is used for include:

A

Methylation/actelylation of histones and where located in genome. Where polymerase or helicase loads for origin of replication. And transcription factors

30
Q

Rh neg is recessive or dominant?

A

Recessive

31
Q

Antigen D is:

A

Rh factor

32
Q

What is agglutination?

A

Aggregate of antibody and antigens present on on cells = sticks to more than one and creates clumps of cells and antibodies

33
Q

What does anti-D prevent?

A

Agglutination. Given to Rh mothers to prevent the production of maternal antibodies against fetal Rho/D antigen

34
Q

The MAb: adalimumab/Humira/Amjevita targets what to prevent?

A

Targets TNF-alpha to prevent inflammation cascade

35
Q

The MAb: Ixekizumab/Taltz targest what to prevent?

A

Targets IL-171A prevents inflammation by inhibiting binding. Humanized IgG4

36
Q

The MAb: Pembroizumab/Keytruda targets what to release?

A

Targets PD-1 to release cell cycle repression of T-cells

37
Q

The MAb: Ustekinumab/Stelara targets what to prevent?

A

Targets IL-17A to prevent inflammation. Human IgG1

Molecular (6 decks)

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