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Y2S1 302 > 13: Antibody drugs > Flashcards

13: Antibody drugs Flashcards

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1
Q

what are polyclonal antibodies

A

Aby secreted by many different clones of B cells against different antigenic epitopes

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2
Q

polyclonal antisera is generated by

A

inoculation of a suitable animal host species with multivalent antigent

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3
Q

what are phylogenetic considerations

A

considering the relationship between the donor Ag species and the recipient Aby producing species

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4
Q

when producing polyclonal antibodies, the more distant the species the _____ the immune response, and the ____ Aby is generated when exposed to donor Ag

A

greater

more

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5
Q

_________________ animals may be required to minimize infectious disease cross over to recipient of Aby

A

specified pathogen free animals

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6
Q

describe how polyclonal antibodies are made

A

disease Ag presented to animal
animal produces Aby from lots of different B cell clones
different types of Aby for this one Ag now put back in infected human

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7
Q

in goat anti-mouse IgG, what is the host species and the antigen presenting species

A

host = goat

antigen presenting = mouse

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8
Q

in horse tetanus antitoxin, what is the host species, what does it neutralize

A

horse = host

neutralizes tetanus toxin

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9
Q

what 2 kinds of cells do you need to make hybridomas

A

immortal myeloma cells + mouse splenocytes

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10
Q

which cells are deficient for enzymes that recycle nucleotides for DNA synthesis in hybridomas

A

myelomas

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11
Q

how many times on average are mouse immunized

A

3

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12
Q

how to screen that the hybridoma is secreting the appropriate Aby

A

ELISA
flow through cytometry
western blotting

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13
Q

list the 4 steps of hybridoma production

A
  1. immunization
  2. fusion
  3. screening
  4. subcloning
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14
Q

bacteriophages are

A

viral molecules that infect bacteria

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15
Q

phage display technology generates Aby like fragments called

A

single chain fraction variable (scFv)

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16
Q

describe the process of phage display

A

coat antigen on plate
add phage library
wash and see which sticks- repeat 3x
infect e.coli with it to amplify + rescreen or ELISA
ELISA to find antigen binding clones
IPTG induced large scale expression of identified clone

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17
Q

what antibody is found in egg yolks

A

IgY

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18
Q

how to make your desired antibody in an egg

A

immunize chicken with antigen 2x at ~21 weeks with pathogens, toxins, biomarkers, allergens, etc and it’ll lay an egg for you with the specific IgY

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19
Q

applications of chicken egg Aby

A

diagnostic reagents

biomedical applications

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20
Q

how to get biotech products from natural sources

A

extraction + protein separation and fractionation

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21
Q

describe recombinant techniques for biotech products

A

recombination of DNA genes + vector genes which are then introduced to host cells to secrete the coded proteins

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22
Q

what are recombinant techniques used for

to manufacture what kind of drugs/ ligands?

A

to manufacture larger protein ligands and cytokine drugs

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23
Q

describe chemical synthesis for biotech products

A

amino acids attached step by step to a solid support

- there is a solution phase and a solid phase

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24
Q 
peptides = \_\_ AA
proteins = \_\_\_ AA
A 
peptides = 2-40AA
proteins = >40AA
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25
what are semisynthetic approaches to making biotech products
chemical mod of natural peptides
26
issues with absorption of peptide drugs
poor absorption across membranes + low oral bioavailability
27
peptide drugs typically have a ___ Vd and ___ half life
low, low
28
peptide drugs are prone to ______ and/or ______degredation
aggregation | chemical degradation
29
why do peptide drugs have low Vd
large size
30
molecules larger than __ can't pass through glomerular membrane for urinary excretion
70kDa
31
protein _____ _____ increases when GFR decreases = greater diffusion across barrier
sieving coefficient
32
chemical degradation breaks ____ bonds resulting in
covalent | chemical structural changes
33
physical biotech drug structure stability depends on
protein secondary, tertiary, or quaternary noncovalent interactions
34
which degredation breaks noncovalent bonds, chemical or physical
physical
35
oxidation is ____ degradation, aggregation is _____ degradation
chemical | physical
36
denaturation is the loss of ___ or ___ structure, resulting in irreversible or reversible changes
secondary or tertiary
37
what sterilization methods should not be used on proteins
heat, gas, ionizing radiation
38
biologics should be assembled under ___ conditions and a final _________ should be done
aseptic | final filtration sterilization (pre filter, then 0.22um filter)
39
T or F: excipients used for biologics can not be sterilized by heat either
F- equipment and excipients sterilized separately from protein API, then combined
40
pyrogen removal mainly targets
endotoxin (LPS) from gram - bacteria
41
how are pyrogens removed
ion exchange chromatography reverse osmosis generated water for injection dialysis, charcoal, oxidation, dry heating
42
what are some excipients used in biotech drugs for parenteral formulations
solubility enhancers anti-adsorption/ aggregation agents buffers
43
what is the shelf life of aqueous biologics
<2 years
44
stability of aq biologics depend on
pH, ionic strength, temperature, excipient stabilizers
45
why do we freeze dry biologics
because the presence of water promotes physiochemical degradation = can extend shelf life to >2 years without it
46
what excipients does freeze drying require
bulking agents collapse temperature modifier lyoprotectant
47
dried, compact biologic tablets are mostly used for ______ and are applied ______ through compressed air powered rifles
used for animals by vets | applied subdermally
48
what kind of oral biologic formulations seem to show improved GI uptake
lipidized
49
what are some limitations to IM and SC
may result in metabolic inactivation before hitting systemic circ bioavailability limited by circulation to specific area variable residence time and disposition
50
T or F: the transdermal route avoids first pass
T
51
what is the primary challenge of the transdermal route
proteins and peptides do not penetrate the stratum corneum well
52
how can we increase ability of drugs to penetrate transdermally
penetration enhancers- microneedle patches, sonophoresis, iontophoresis
53
what is a noninvasive approach that avoids first pass
nasal administration
54
what is a barrier for bioavailability in nasal admin
large molecular weighr
55
why is buccal sometimes preferred over GIT
less protease and acid degradation
56
what is a problem with pulmonary admin route
lung irritation and induction of immune response
57
what are biotech products
when living organisms are involved in the production or modification of a product
58
what are biologic response modulators
immunomodulators that target disease causing mechanism
59
what are biosimilars
a biological product that is highly similar to a reference product and show no clinically meaningful difference from reference product in terms of safety, purity, and potency
60
how do biologic drugs differ from small molecule drugs
structural complexity | manufacturing techniques
61
what are some patient related factors that can influence biologic ADRs
genetic predisposition to produce neutralizing antibodies expression of the endogenous eq of the product comorbid conditions
62
T or F: immunogenisity from biologics are usually mild
F- range from irrelevant to lethal- unpredictable
63
excipients are added in order to increase biotech drug ____________
stability and preservation maintain tonicity facilitate drug delivery avoid immunogenicity
64
protein based therapeutics may cause an ____ response leading to AEs, or drug ____ upon repeated exposure
immunogenic | drug clearance
65
excipient categories include
``` pH modifier tonicity agents bulking agents wetting and//or solubilizing agent antioxidants antimicrobial preservative chelating and/ or complexing agents ```
66
on average, how many excipients does a biological drug product contain?
4.45
67
T or F: small molecule oral medications contain less excipients on average compared to biological drug products
F- bio = 4.45, molecule = 8.8
68
what are the 4 most common excipients in biotech drugs
water sodium chloride polysorbate 80 sucrose
69
how can clinicians provide further investigational evidence of ADR to medications
skin prick testing + other diagnostics
70
describe BRM therapy
a type of treatment that mobilizes the body's immune system to fight cancer
71
describe the cytokine scavenging Aby drug in terms of bone resorption
binds cytokines so that it can't release signals and stimulate osteoclasts to breakdown bone
72
what is chimeric mAb
an antibody that has components from both mouse and human
73
what is the prefix associated with mouse Aby
xi
74
how to make humanized mAb
graft CDRs from a mouse antibody onto a human variable region framework
75
what prefix is used for humanized Aby
zuman
76
fully human Aby prefix
umab
77
fully human antibodies have been achieved using ____ + _______
phage display and mammalian cell culture
78
infliximab is a __ aby and binds and neutralizes ___
mouse chimeric Aby | binds TNF-alpha
79
adalimumab is a ____ aby that binds _
fully human antibody | TNFalpha
80
what are drug delivery systems
specially designed formulations or devices that can deliver the drug cargo exactly where it needs to be at a controlled rate + minimize systemic exposure
81
list some reasons why DDS are needed
increases ease of administration increase deposition at specific site = decreased AE and lower dose needed resolve formulation issues
82
T or F: companies may get FDA approval for a drug tablet, they can then make this drug into a solution and sell it
F- only what form is approved can be made
83
what are the 5 features of an ideal DDS
``` ease of administration able to control the release of API able to deliver API at site needed low immunogenic potential/ hypersensitivity nontoxic ```
84
what are some DDS delivery vehicles
liposomes, polymers, nanoparticles
85
what are the 6 factors affecting selection of route of administration
``` drug characteristics ease of administration mechanism of action onset of action + duration quantity of drug to be administered ```
86
T or F: DDS can alter the physiochemical and PK properties of API
T
87
what is naked antibody targeting
unmodified antibodies used as BRMs, cytokine scavengers, receptor agonists/ antagonists, etc
88
what is rituximab target? is it mono or polyclonal
monoclonal Aby that targets CD20+ B cells in RA
89
antibody drug conjugates are
antibodies with targets that deliver drug cargo
90
how to make antibody- drug conjugates
must use immunoconjugation strategies- only a small number of protein FG serve as practical targets for bioconjugation
91
what protein FG can be targets for bioconjugation to add drugs onto Aby
primary amines carboxyls sulfhydryls carbonyls
92
what are theranostics in Aby
antibody isotope conjugates for imaging and therapy
93
fusion proteins created through joining of 2 or more genes that originally coded for separate proteins describes
antibody fusion protein constructs
94
how do antibody fusion protein constructs work
newly bound antibody and protein can act as decoy receptors for inflammatory proteins = bind and don't activate complement to decrease inflammation
95
abatacept uses __________ technology
antibody- fusion protein
96
how are immunoliposomes made
vortex lipids with ethanol = forms unilamellar liposomes by extrusion through stacked polycarbonate membranes post insertional technique to transfer targeting peptide into bilayer of liposome
97
at what temperature are immunoliposomes made
stir at 37C for 5hrs, then 19h at room temp
98
liposome release can be triggered by
``` pH decrease external alternating magnetic field hyperthermia light ultrasound ```
99
what are quadromas
bispecific/ bifunctional Aby for drug delivery targeting
100
what are quadromas useful for
crosslinking 2 distinct targets | rapid immunodiagnostic kits
101
what is the current chemical fusogen of choice for hybridomas
PEG
102
T or F: immunization of mice to produce hybridomas require adjuvants
T
103
viable hybridomas are ___ medium selected
HAT- a medium without functional thymidine analogues = only cells with active TK enzymes can survive
104
do peptides and proteins penetrate the stratum corneum of the skin
no
105
which of the following may result in metabolic inactivation before reaching systemic circulation + are limited by circulation to specific area 1. IV 2. IM 3. SC 4. Nasal 5. buccal
2, 3
106
which show greater heterogeneity, biologic drug products or small molecule drugs
biologic- protein folding + bonds
107
which antibody therapy is more likely to have to be injected IV, dufliximab or yuzumumab?
dufliximab because it's a mouse chimeric Aby = more likely to be recognized by immune system than yuzumumab a human antibody = must be injected IV instead of SQ

Y2S1 302 (10 decks)

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