Genomics Final

What are the steps of biopharmaceutical development?

"1. mRNA isolated from human cells, 2. double stranded cDNA produced 3. cDNA ligated into a plasmid (vector) 4. Plasmid transfected into host cells 5.Host cells produce human protein (large fermenter), 6.Protein is isolated from host cells (harvest, purification, isolation)"

Exondys 51 (Eteplirsen) mechanism of action

(Eteplirsen) binds to dystrophin primary transcript exon 51 with stop codon mutation spliced out shorter dystrophin protein expressed

The EPO cDNA was discovered and cloned into a mammalian vectors & the vector was transfected into mammalian host cells. What are those cells called?

- Chinese hamster ovary (CHO) cells

As delivery agents, Mab specificity should:

- Reduce the dose of molecule to be delivered - Reduce side effects (non-specific killing) -active at the site after delivery

What are the requirements for Mabs as delivery agents ?

- bond that conjugates the antibody to the molecule should be stable -must retain ability to bind to target

What's is the advantages of Mab therapy?

- no cross resistance w/ traditional cytotoxics/ radiotherapy - tumor cells resistant to radiotherapy or traditional are not resistant to lungs - solid tumor: majority of cancer cells aren't dividing -limits the action of cytotoxic drugs

What are the mechanisms of resistance (challenges) in Mab ?

-Immune reactions can still occur against Chimeric or humanized antibodies in some patients (monitor closely) -Soluble tumor antigen shed by tumor -Tumor heterogeneity (Not all malignant cells within a single tumor express the same antigen)

How does Anticancer Mabs work?

-Interference w/ receptor activation -Interference w/ intracellular signaling after internalization -effector function: antibody interactions with immune cells via Fc receptors • Antibody-dependent cell-mediated cytotoxicity (ADCC)

What are the clinical signs and symptoms of Pompe disease in infants ?

-cardiomyopathy -severe generalized hypotonia -enlarged tongue -hepatomegaly -feeding difficulties/thrive failure -respiratory failure

In the gene therapy treatment for skin condition epidermolysis bullosa, What happens after the 1st step?

-cells infect with a lentiviral vector with a functional laminin B3(LAMB3) cDNA -cells were grown into patches of transgenic epidermal grafts -3 graft operations replaced of the patients skin with the grafts (holo/mero/paraclone)

Mabs recognize specific targets to

-directly interfere with target molecules/cells - to deliver different molecules to targets by attaching molecules to the Fc region

What is the appropriate medication administration instructions for Nexviazyme ?

-infuse using separate line -use low protein-binding filter - infuse per instructions -must be completed by 9 hrs of dilution/ 9 hrs of fridge removal

What is the appropriate medication preparation, administration, and monitoring instructions for Lumizyme®/Myozyme® ?

-prepare: avoid foaming, avoid shaking, mix in appropriate volume (take air out bag), and protect from light -adminstration: infuse over 4 hrs, gradually increase infusion rate -monitor: vital signs

What is the appropriate medication preparation instructions for Nexviazyme ?

-room temperature prior to reconstitution, -avoid foaming -view if discoloration / particles -dilute to conc. large volume -administer immediately

What are the clinical signs and symptoms of Pompe disease in juvenilles and adults (late onset) ?

-skeletal myopathy -respiratory failure -delayed gross motor development -limb girdle progressive weakness

Which of the following are solutions to RNA therapies challenges ?

-synthesizing RNA molecules w/ modified backbones ex: replacing phosphates w/ thioate linkages btwn sugars morpholino rings instead of ribose -formulations thats enhance stability ex: nanoparticle systems used in mRNA vaccines

Which of the following are challenges of gene therapy?

-the body sees these treatments as "foreign" and attacks - patients with antibodies excluded due to prior exposure - treatments can lead to cancer if it integrates in the wrong part of the genome -ONE time treatments (immune rxns prevent repeated doses)

Which pharmacotherapy management strategies should be used for supportive care and treatment of Pompe disease ?

-vaccines, albuterol, mucomyst for pulmonary system -enalapril for cardiovascular -ergocalciferol (Vitamin D2), polyethene glycol, and formula for feeds

steps of immunofluorescence

1) samples placed on microscope slides 2) antibody added and binds to target protein 3) second antibody with fluorescent tag (*no enzyme*) recognizes 1st antibody 4) visualized with fluorescent microscope

Steps of epoetin production in host cells

1. Host cells transcribe the EPO cDNA into mRNA 2. Translate mRNA into protein 3. The EPO protein is glycosylated (N- and O-linked, post-translational mods.) 4. Glycosylated EPO is secreted by the host cells • 5. EPO is harvested, purified, and formulated

Match a GENE with the disease. 1. Epidermolysis bullosa 2. Retinal Disease 3. Spinal Muscular Atrophy

1. LAMB3 2. RPE65 3. SMN1

Match the following RNA therapeutics to it's target. 1. Onpattro (patisiran) 2. Spinraza (nusinersin) 3. Exondys 51 (eteplirsen)

1. TTR mRNA 2. SMN2 primary transcript 3. dystrophin primary transcript

List the steps of Immunohistochemistry (IHC)

1. Tissue sample is isolated and treated 2. Sample is sliced into thin sections and placed on microscope slides 3. Antibody is added & will specifically bind to target protein 4. secondary antibody with enzyme added 5. Protein is visualized

Why target RNA molecules ?

1. inhibit message from being translated 2. cause degradation of the message 3. cause alteration of the message (improve it's translation)

Initial biotech production of human insulin steps

1. mRNA isolated from pancreatic islet cells 2. double stranded cDNA produced 3. cDNA ligated into a bacterial plasmid 4. plasmid transfected into bacteria 5. bacteria produced proinsulin 6. proinsulin purified from bacteria and converted to insulin

Hybridomas are grown to ____1___ Mab of single specificity. Mabs are purified by _____2____

1. mass produce 2. affinity chromatography

List the steps of a western blot

1.Gel electrophoresis seperate proteins 2. proteins are transferred (blotted) to a membrane 3. Primary antibody is added and will specifically bind to target protein 4. secondary antibody binds to the primary antibody 5. Band is visualized

Which of the following contain non-native amino acid changes to accelerate or prolong duration of action?

2nd generation Human Insulins

Whats the specific activity of Alglucosidase alpha (Myozyme)

3 to 5 U/mg one unit is defined as that amount of activity that results in the hydrolysis of 1micromole of synthetic substrate per min

What are the advantages and disadvantages of viral based approaches ?

Advantages:viruses good @ infecting cells Disadvantages: -where is DNA integration in genome -pre/existing immunity/immune reaction to the virus

What is Papain protease?

An enzyme that cleaves the IgG antibody molecule above the disulfide bond

What does monoclonal antibodies from mouse and other animals cause following repeated doses ?

B-cells cause severe immune reactions (Anaphylactic)

What cancer is related to over-expression of HER2(human epidermal growth factor receptor 2) ?

BREAST cancer

Which of the following have aggressive defense mechanisms against any foreign DNA/ RNA ?

Bodies and Cells

Where are the Peg groups added to on Certolizumumab (Cimzia ™ ) ?

C-terminus end of Fab where absent Fc region would be

Which cancer immunotherapy takes autologous T cells from blood, genetically modify it , stimulate it, then reintroduce it into the patient to attack the cancer?

CAR-T cell therapy (Chimeric Antigen Receptor)

Which region is the variable region of Fab that directly binds to antigen on N-terminus ?

CDR (complementary determining region)

What is used as a diagnostic test for HER2 gene amplification and uses a label on a DNA probe to allow detection by IHC?

CISH (Chromogenic In Situ Hybridization)

Pembrolizumab (Keytruda™) and nivolumab (Opdivo™) are immunotherapy "checkpoint blockade" anti-cancer antibodies How does this work?

Cancer cells w/ dMMR = 1,000 point mutations resulting proteins to become MANAs checkpoint blockade facilitates immune recognition of the cancer cell as "non self"

Clinical features of Pompe disease

Cardiorespiratory failure, causes of death usually before 4

Which drugs bind to non-overlapping epitopes of the spike protein RBD (receptor binding domain) of CoV-2 preventing the spike protein to bind to human ACE-2 receptors?

Casirivimab & imdevimab (Mab therapeutics that target SARS-CoV-2)

Which drug is a humanized Fab with 2 PEG groups attached to the C-terminus end and the absence of the Fc region reduce antibody-dependent cellular cytotoxicity ?

Certolizumumab (Cimzia ™ )

Which of the following is an cytokine example that stimulates cell growth, One example: granulocyte-CSF (G-CSF), and PEG group added to improve properties?

Colony Stimulating Factors (CSFs)

Which condition is where stop codon mutations prevent expression of protein w/ C-terminus?

Duchenne MD

Which immunoassay uses antibodies to detect proteins by color change or fluorenscence?

ELISA (enzyme-linked immunosorbent assay)

What are all the different types of immunoassays ?

ELISA, Westerns, IHC and immunofluorescence are all different examples of immunoassays. They use antibodies to detect proteins.

glycosylation occurs in

ER and Golgi • Complex and terminal glycosylation occurs in the Golgi some occurs in ER

was first used to treat anemia associated with chronic renal failure, has also been tried in the treatment of anemia that results from many conditions, and restricted use due to cancer-associated anemia?

Epoetin Alfa (Epogen®, Procrit®)

Which of the following is a natural hormone produced by the kidneys to stimulate red blood cell growth and stimulate erythocyte burst formings units (BPU-E) to divide and form mature erythocytes ?

Erythropoietin (EPO)

Whats the difference between the other Mab therapeutics that target TNF-a and Cimzia ?

Fab (antigen-binding fragment)

What's the importance of adding 2 PEG groups on the Fab fragment of Certolizumumab (Cimzia ™ )?

Fab fragments have a much shorter half-life than full length antibodies...so the peg groups will increase half life

Which region of antibody allows for glycosylation?

Fc region (constant part of heavy chain)

Where does drugs, enzymes, fluorescent, and radioactive agents bind to antibodies ?

Fc region of the antibody

Which of the following is obtained from the bacterial fermentation of a strain of Escherichia coli transformed with a genetically engineered plasmid containing the human G-CSF gene?

Filgrastim

Which of the following require delivering nucleic acids into cells ?

Gene Therapy and RNA therapies

In insulin sequence comparison, pig and human insulin is different by only one amino acid in B-chain

Human amino acid = Thr Pork= Ser

recombinant human insulins

Humulin (Produced in E. Coli) Novolin R ( Produced in S.cerevisiae)

Which antibodies induce pro-inflammatory effector functions ?

IgG1 Mabs

What does the corona virus ELISA test collect ?

IgM/IgG from blood sample

What is used to visualize protein directly in tissue biopsy, histology or sample?

Immunohistochemistry (IHC)

In the gene therapy treatment for skin condition epidermolysis bullosa discussed in class, which of the following best describes the first step in the process for a patient?

Isolate skin cells from an unaffected area of the patient.

What is the normal mechanism of action for normal PDL- receptors ?

It inhibits T cell activation and acts as a natural "dampener" to prevent overactive immune responses Tumor cell express PD-L1 to avoid immune attack from T cells

Which drug is used for elevated HER2 expression by IHC to FIGHT against Herceptin (Trastuzumab)-resistant tumors ?

KADCYLA® (trastuzumab emtansine)

Which insulin is produced in E. coli, Glycine replaces ASN ( asparagine) at position 21, 2 C-terminal Arginine added, microprecipitates upon injection, Decreased solubility, Slower acting ?

Lantus

What are mice genetically engineered with in next generation humanized Mabs to not express murine globins ?

MEGAbase-sized YACs(yeast artificial chromosome) carrying portions of human immunoglobulin

Which region of the antibody determines if it can bind to a particular antigen?

N-terminal region of Fab (antigen-binding fragment)

Which insulin is produced in S. cerevisiae, Aspartate replaces proline at position 28 on beta chain, Decreased oligomerization, and is Rapid acting?

Novolog

Nusinersen (Spinraza®) was approved by the FDA for the treatment of spinal muscular atrophy (SMA), which is caused by the loss of the Survival Motor Neuron (SMN) protein. Which of the following best describes the mechanism of action ?

Nusinersen binds to the SMN primary transcript, preventing an exon from being spliced out.

Which of the following are viruses engineered to selectively infect and replicate in tumor cells, causing tumor cell lysis (death) and an immune response?

Oncolytic viruses

Pertuzumab (perjeta) is an antibody that targets HER2 differently from trastuzumab (herceptin). Which of the following best describes how Pertuzumab targets HER2?

Perjeta targets the extracellular dimerization domain of HER2

What is the one source of severe inflammatory syndrome in COVID ?

Pro-inflammatory IgG1 immunoglobulin levels higher than other other IgGs

Which of the following cause retinal disease, vision deterioration and progression to blindness ?

RPE65 gene mutations

Which drug have shorter half life because it only have a the Fab region of its antibody and is missing Fc region ?

Ranibizumab (Lucentis)

What disease is caused by elevated circulating levels of TNF-a ?

Rheumatoid Arthritis (R.A.) and other autoimmune inflammatory diseases

There are 4 types of Spinal Muscular Atrophy (SMA) TYPE I, II, III, IV (Severity) Severity is dependent on residual _____ expression.

SMN2

What was originally identified as a molecule that can kill cancer cells and then was found out to be a pro- inflammatory cytokine and plays a role in immune responses?

TNF-a (Tumor necrosis factor)

In humans, insulin gene introns are spliced out

TRUE

What are the differences between chimeric and humanized MoAbs?

The chimerics were developed by recombinant DNA cloning of the DNA sequences of mouse-derived B cells fused with myeloma cells (hybridomas) that produced those antibodies with only a little residual mouse sequence. Humanized ones were generated by challenging transgenic mice with human immunoglobulin genes, then isolating B cells to create hybridomas. So, the humanized ones really don't have any mouse residual Fc or Fab sequence because of the humanized IgG genes in the mice.

What is the difference between FISH & CISH?

The difference between FISH and CISH is that F stands for fluorescence, so fluorescent DNA probe. C stands for chromogenic, which in this case utilizes two antibodies and similar to secondary ELISA. Primary antibody in CISH is detecting a chemical tag, abbreviated DIG, covalently attached to the DNA probe, which binds to DNA. Then the secondary antibody with enzyme attached detects the Fc region of the primary antibody.

Which of the following is one similarity between a sandwich ELISA and an indirect ELISA?

The enzyme is covalently attached to the secondary antibody in both assays.

In gene therapy, "ex vivo" means...

The introduction of the genetic manipulation begins outside the body of the patient.

In gene therapy, "in vivo" means...

The introduction of the genetic manipulation begins within the body of the patient.

How to produce pegfligastrim from Fligrastim?

To produce pegfilgrastim, a 20 kD monomethoxypolyethylene glycol molecule is covalently bound to the N-terminal methionyl residue of Filgrastim

Which condition is an autoimmune destruction of the pancreatic B-cells that produce insulin ?

Type 1 diabetes

What is over expressed in colorectal cancer ?

VEGF (vascular endothelial growth factor)

What are the targets of Aflibercept (Eylea)?

VEGF-A, VEGF-B, and placental growth

Which of the following utilizes the ability of viruses to infect cells ( deliver DNA/RNA) after inactivating the ability of the viruses to replicate and propagate ?

Viral based approaches

Compared to mammalian cells, bacteria or yeast are _____[X] and ______[Y] to grow.

X= less expensive Y= easier

Which drugs have a recombinant fusion protein of VEGF receptor binding domains and an Fc region?

Zaltrap ® (ziv -aflibercept) and Eylea ®(aflibercept)

Which of the following treats Spinal Muscular Atrophy?

ZolgenSMA

Within the bacteria or yeast host cells, the proinsulin cDNA must be properly [1] and [2] to produce the proinsulin protein.

[1] transcribed and [2] translated

Which of the following are measures for determining successful protein purification?

a. Good yield b. Specific activity maintained (enzymes) c. Structural integrity maintained

Which of the following modifications is used to improve the stability of Exondys 51 (Eteplirsen)? a. Morpholino ring in place of ribose b. Addition of a methoxyethyl group to RNA bases c. Lipid nanoparticle delivery system d. Thioate linkages within RNA backbone

a. Morpholino ring in place of ribose

Which of the following RNA therapeutics does NOT bind to an RNA primary transcript? a. Onpattro (Patisiran) b. Spinraza (Nusinersen) c. Exondys 51 (Eteplirsen)

a. Onpattro (Patisiran) primary transcript= before splicing

Which of the following diseases are treated with in vivo therapy? a. Spinal Muscular Atrophy b. Epidermolysis bullosa c. Retinal Disease

a. Spinal Muscular Atrophy c. Retinal Disease

In this example of indirect ELISA, The primary antibody... a. comes from the patient's blood sample if they have previously been exposed to the virus. b. is covalently attached to the plate. c. recognizes the Fab region of the secondary antibody. d. converts a colorless substrate into a colored product.

a. comes from the patient's blood sample if they have previously been exposed to the virus.

In proinsulin sequence, the connecting peptide is removed to release ______.

active insulin

A subset of breast cancers are characterized by overexpression of HER2. This overexpression is caused by ______ of the HER2 gene.

amplification

What is a protein synthesized by an organism in response to the presence of a foreign substance (antigen)?

antibody

HER2 amplification is test twice in cancer cells. What are the two test ?

antibody test IHC (detects Proteins) gene amplification test FISH (detects DNA)

As shown in the video about the hCG home pregnancy test, the control (C) strip contains... a. A monoclonal antibody that recognizes hCG. b. A monoclonal antibody that recognizes the mouse antibody. c. Polyclonal antibodies that recognize hCG. d. Polyclonal antibodies that recognize the mouse antibody.

b. A monoclonal antibody that recognizes the mouse antibody.

Which of the following modifications is used to improve the stability of Spinraza? a. Morpholino ring in place of ribose b. Addition of a methoxyethyl group to RNA bases c. Lipid nanoparticle delivery system d. Thioate linkages within RNA backbone

b. Addition of a methoxyethyl group to RNA bases d. Thioate linkages within RNA backbone

Which of the following diseases are treated with a holoclone? a. Spinal Muscular Atrophy b. Epidermolysis bullosa c. Retinal Disease

b. Epidermolysis bullosa

Which of the following statements best describes the mechanism of action for Atezolizumab (Tecentriq™)? a. It binds to the HER2 ligand, is internalized into cancer cells, and releases an anti-mitotic drug. b. It binds to the PD-1 ligand on cancer cells, which relieves the inhibition of T cells. c. It binds to the PD-1 ligand on T cells, shielding T cells from being attacked by cancer cells. d. It binds to the HER2 ligand on T cells, which shields T cells from being attacked by cancer cells.

b. It binds to the PD-1 ligand on cancer cells, which relieves the inhibition of T cells.

Which of the following is NOT an advantage of bacterial/yeast sources for biopharmaceuticals?. a. Higher yield b. Limited post-translational modifications c. Easier to grow d. Cheaper

b. Limited post-translational modification

Fluvoxamine metabolism is affected by genotype and environmental interactions, namely smoking. Which of the following statements best represents the influence of smoking on fluvoxamine metabolism? a. Tobacco smoking suppresses the expression of CYP2D6, which increases the plasma concentration of fluvoxamine. b. Tobacco smoking increases the expression of CYP2D6, which decreases the plasma concentration of fluvoxamine. c. Tobacco smoking suppresses the expression of CYP2D6, which decreases the plasma concentration of fluvoxamine. d. Tobacco smoking increases the expression of CYP2D6, which increases the plasma concentration of fluvoxamine.

b. Tobacco smoking increases the expression of CYP2D6, which decreases the plasma concentration of fluvoxamine.

immunofluorescence is a. Used to visualize protein directly in a tissue biopsy or sample b. Used to visualize protein within a cell

b. Used to visualize protein within a cell

In this example of sandwhich ELISA, The primary antibody... a. comes from the patient's blood sample if they have previously been exposed to the virus. b. is covalently attached to the plate. c. recognizes the Fab region of the secondary antibody. d. converts a colorless substrate into a colored product.

b. is covalently attached to the plate.

In principle, antisense oligonucleotides work as therapeutics with a high degree of specificity because... a. they are identical to an endogenous RNA b. they are complementary to an endogenous RNA c. they are identical to an endogenous protein d. they bind to an endogenous protein, causing its degradation.

b. they are complementary to an endogenous RNA

proinsulin protein synthesis is in _____

bacteria/yeast because bacteria do not splice out introns

bacterial yeast/mammalian sources have HIGHER yelid

bacterial yeast

Onpattro (patisiran) Mechanism of action

bind to TTR mRNA TTR mRNA complex recognized by RISC TTR mRNA cleaved by RISC which prevents translation into protein

generic biopharmaceuticals

biosimilars

Which immunoglobulin type is the major antibody of secondary (adaptive) immune responses? a. IgM b. IgD c. IgG d. IgA

c. IgG

Which of the following modifications is used to improve the stability of Onpattro (patisiran)? a. Morpholino ring in place of ribose b. Addition of a methoxyethyl group to RNA bases c. Lipid nanoparticle delivery system d. Thioate linkages within RNA backbone

c. Lipid nanoparticle delivery system

Which of the following best describes the mechanism of action of acyclovir (Zovirax)? a. Phosphorylated by viral thymidine kinase, incorporated into DNA by viral polymerases and blocks translation. b. Phosphorylated by human thymidine kinase, incorporated into DNA by viral polymerases and blocks transcription. c. Phosphorylated by viral thymidine kinase, incorporated into DNA by viral polymerases and blocks replication. d. Phosphorylated by viral thymidine kinase, incorporated into DNA by human proteases and promotes replication.

c. Phosphorylated by viral thymidine kinase, incorporated into DNA by viral polymerases and blocks replication.

As shown in the video about the hCG home pregnancy test, the test (T) strip contains... a. A monoclonal antibody that recognizes hCG. b. A monoclonal antibody that recognizes the mouse antibody. c. Polyclonal antibodies that recognize hCG. d. Polyclonal antibodies that recognize the mouse antibody.

c. Polyclonal antibodies that recognize hCG.

Which one enzyme below is known in its gene to contain natural human polymorphisms that affect the metabolism of fluoropyrimidine drugs such as Fluorouracil? a. adenosine deaminase b. DNA helicase c. dihydropyrimidine dehydrogenase d. inosine dehydrogenase

c. dihydropyrimidine dehydrogenase

In this example of sandwhich ELISA, The antigen a. comes from the patient's blood sample if they have previously been exposed to the virus. b. is covalently attached to the plate. c. recognizes the Fab region of the primary antibody. d. converts a colorless substrate into a colored product.

c. recognizes the Fab region of the primary antibody.

What is the name of the enzyme that converts mRNA into a cDNA? a. DNA ligase b. RNA ligase c. reverse transcriptase d. DNA helicase

c. reverse transcriptase

Biopharmaceutical versions of Epoetin were originally produced in mammalian CHO cells because they...

can carry out complex post-translational modification *advantages

What is the function of α-1,4-glucosidase (lysosomal)?

catalyze the break down of glycogen

What are side effects of radiation and chemotherapy ?

cause myelosuppression (decrease neutrophils & platelets), mucositis (mouth & GI ulcers), and nausea/vomiting

What are side effects of Mab therapy ?

causes fever and chills with the infusion

What is the appropriate monitoring instructions for pompe treatment ?

check for IgG antibodies, liver function test (LFTs), vital signs, and volume overload

What are toxins from killed bacteria used in cancer chemotherapy to kill surface tumors?

coley toxins

In a previous year, a student asked about an application of Indirect ELISA. How would a test start with the antigen coated in the well? A clinical example is a test to determine if someone has been exposed to Herpes Simplex Virus. The test plate has herpes simplex viral particles covalently fixed in the wells. A blood sample from patients to be tested are added to wells to begin the assay. In this example of indirect ELISA, The primary antibody...

comes from the patient's blood sample if they have previously been exposed to the virus.

what are the limitations of protein biopharmaceuticals?

cost, injections, degradation (denature in stomach), instability

______ biopharmaceuiticals are polypeptides that act in inflammation and immune responses

cytokines

Multiple anti-coronavirus Mabs have received EUA as experimental therapeutics against COVID. Which of the following best describes the mechanism of action for these antibodies? a. They recognize different antigens corresponding to different proteins of the SARS-CoV-2 virus. b. They recognize different antigens corresponding to the ACE2 protein of human cells. c. They recognize the same epitope on the spike protein of the SARS-CoV-2 virus. d. They recognize different epitopes corresponding to the spike protein of the SARS-CoV-2 virus.

d. They recognize different epitopes corresponding to the spike protein of the SARS-CoV-2 virus.

Which of the following bests describes Immunohistochemistry (IHC)? a. Uses PCR to detect DNA by a change in size. b. Uses Reverse Transcriptase (RT)-PCR to detect mRNA by a color change. c. Uses PCR to detect proteins by a color change. d. Uses antibodies to detect proteins by a color change. e. Uses gel electrophoresis to detect proteins by a change in size.

d. Uses antibodies to detect proteins by a color change.

In ELISA, the enzyme... a. comes from the patient's blood sample if they have previously been exposed to the virus. b. is covalently attached to the plate. c. recognizes the Fab region of the secondary antibody. d. converts a colorless substrate into a colored product.

d. converts a colorless substrate into a colored product.

What is induced when TNF-a binds to TNF receptors on cells?

death and inflammation

In the structure of the active insulin protein, what holds the A and B chains together?

disulfide bonds

MoAb anticancer therapeutics are antibodies raised against specific proteins in certain cancers. These MoAbs are ONLY used in those cancers in which the targeT protein levels are ________, relative to normal tissue.

elevated

We use gene therapy because the _ gene is missing or mutated.

endogenous

What is the specific portion of the antigen that is recognized by antibody?

epitope

Gene therapy is the introduction of _____ genetic material to correct or modify the function of a cell.

exogenous

Eteplirsen splices out ___________ which allows for the production of functional dystrophin protein.

exon 51

8-13% of Duchenne MD patients have truncating (shortened in duration) mutations in ______

exon 51 *mutations can also occur exon 53

RNA are chemically unstable and also rapidly degraded by _________.

exonucleases

What are the similar challenges of DNA and RNA therapies?

exonucleases digest DNA and RNA

What is a mechanism of resistance to Mabs regarding PD-1 receptors ?

exosomal PD-L1 contributes to immunosuppresion and is associated with anti-PD-1 response, T cell eventually binds to exosome bond PD-L1 produced by tumor cell & cause resistance to drug because tumor cell was not targeted

After purification, formulation, and reonstitution the protein MUST be correctly ______ and catalytically active/inactive

folded, active

How many red dots needs to be represented of HER2 gene amplification detected by FISH (fluorescence in situ hybridization) ?

greater than 2 red dots

Which of the following is an autosomal dominant inheritance, TTR protein is normally a tetramer, and mutation cause misfolded TTR and aggregation into amyloid deposits ?

hATTR amyloidosis

Disulfide bonds hold ______ in the antibody structure.

heavy and light chains

disadvantages of mammalian sources

higher cost care required for growth lower yields

Which of the following are undifferentiated skin cells that have self-renewal capacity (ex: stem cells) ?

holoclones

______ biopharmaceuiticals are hematopoietic, wound healing, growth

hormone

Severity of muscular dystrophies depend on ..

how much of dystrophin protein is affected

Biosimilars differ from traditional generics because proteins are

huge molecules with intricate structures.

In the production of chimeric and humanized MoAbs, the DNA sequences for the Fab regions are cloned and (ligated) inserted into a cDNA with __________ Fc-encoding sequence

human

Peg addition function

improve properties

What is the downside (warnings) of drugs that target TNF-a?

increase susceptibility to infections due to decreased TNF-a

Bacteria CAN NOT SPLICE OUT ______

introns

With anticancer antibody-drug conjugates, the molecule to be conjugated to the antibody...

is a cytotoxic agent that enhances the activity.

Anemia is an symptom of

kidney failure

What are the adverse effects of pompe disease treatment ?

life-threatining allergic rxn hypersensitivity cardiorespiratory failure infusion reaction

disadvantages of bacterial/yeast sources

limited post-translational modifications

Which of the following causes Spinal Muscular Atrophy?

loss of SMN protein which causes motor neuron loss (SMN1)

Which of the following cause Muscular Dystrophy (MD)?

loss of dystrophin, a protein that stabilize muscle loss cause muscle wasting (weakness)

In the initial development of recombinant human insulin as a biopharmaceutical, the _____ was isolated from human pancreatic islet cells.

mRNA

Arrange steps in order (top to bottom) as they occur in the development of a biopharmaceutical product.

mRNA Isolation from human cells double stranded cDNA is produced ligate cDNA into a plasmid vector transfection of plasma into host cells isolation protein and purification

"in insulin production proinsulin cdna/mRNA isolated from pancreatic islet cells to make _____to Plasmid transfected into bacteria/animal to produce _______, _______ is isolated from bacteria and converted into _______"

mRNA, cdna, bacteria, proinsulin, insulin

Alglucosidase (Myozyme) is produced in mammalian CHO cells. However, terminal sugars must be _______ in order to be taken up by macrophages after injection.

mannose

Which of the following are Antisense Oligonucleotides (ASOs) ?

modified RNA drug that is complementary to an endogenous RNA

What type of antibody recognize a single epitope on a single antigen ?

monoclonal

In a tissue biopsy of Immunohistochemistry (IHC), what does more brown colorization indicate ?

more antigen bound to antibody in sample

Symptoms of Duchenne MD

muscle wasting

B cells from spleen are fused with _______ cells to produce hybridoma cells.

myeloma cells

Which disease cause overexpression of VEGF in the eye and higher mRNA and protein causing major vision loss ?

neovascular macular degeneration (AMD)

The pharmaceutical properties of "biobetters" are improved by ___________ modifications to the natural protein

non-natural

A "Biobetter" is a protein with a _________ modification(s) that improves the pharmaceutical properties of the original protein.

non-natural ex: Peg additions

Luxturna is a ____ gene therapy for each eye.

one-time

What happens after the lipid nanoparticle bound by ApoE and endocytosed into a hepatocyte in Onpattro (patisiran) ?

pH changes after endocytosis release siRNA

______ is used to REMOVE connecting peptide to make proinsulin to insulin

peptidase

What is a mixture of individual antibodies that recognize different epitopes on the same antigen ?

polyclonal antibodies

What is the job of Risdiplam ?

prevents exon 7 skipping of the SMN message

Which of the following is affected by formulation and environmental conditions, including light, temperature, moisture, packaging material, container closure systems, and delivery deliver ?

protein modification and high order structure

HER2 is an example of a

protooncogene, after amplification it becomes an oncogene

In immunoassays, visualization can occur by measuring which of the following?

radioactivity fluorescence color change

In western blots, visualization can occur by measuring which of the following?

radiography or fluorescence

Avastin is just as effective as Lucentis for treating AMD. What has occurred as a result of the recompounding of Avastin for AMD treatments ?

serious eye infections

How to evaluate protein purification sucessfully?

specific activity (enzymes) yield integrity

Hormone/cytokine therapy is very selective. The protein interactions are MORE ___ and have higher ____ than small molecules w/ targets

specific, affinity

What is a Herpes simplex virus-based immunotherapy to treat melanoma ?

talimogene laherparepvec (T-VEC) IMLYGIC

Fab fragments of antibodies can be created from full length antibodies by using which of the following?

the papain protease

The difference between a sandwich ELISA and an indirect ELISA is

the secondary antibody binds to the antigen in the sandwich ELISA.

The news article about gene therapy mentions a "complication of gene therapy" regarding integration of the gene information into the person's genome, which was also listed as a disadvantage in the "challenge" slide. What property of viruses and their DNA integration raises a concern about a potential long-term side effect of viral gene therapy?

the viral gene therapy integration occurs in random sites, which might trigger genes that control tumor development.

Why is alternative splicing used in therapeutics ?

to produce different forms of a protein

RNA interference (RNAi) targets mRNA to prevent the message from being.....

translated

Which of the following is an ex vivo clinical gene therapy example?

treatment for epidermolysis bullosa

Explain the resistance of Mab when soluble tumor antigen shed by tumor

tumor cells may adapt by increasing shed protein which prevent Mab from binding tumor cells. In the blood stream antigens acts as a decoy to prevent the antibody from ever getting to cancer cell

Describe Survival of Motor Neuron (SMN)

two copies of gene (SMN1 &SMN2) one copy is inactive because of alternative splicing (SMN2) -SMN1: naturally expressed in more ppl -SMN2: mutation cause exon 7 to be spliced out

What is the effect of enzyme absence in Pompe disease?

when -1,4 glucosidase is absent, -glycogen increase -lysosomes swell w/ glycogen -lysosomes rupture to form glycogen lakes -myofibril function is disrupted

Generally, protein interactions are more [x] and with higher [y] than a small molecule with their targets.

x= specific; y= affinity

Which of the following is transgenic mice that produce full humanized antibodies ?

xenomouse

What is enzyme is lacking in Pompe disease?

α-1,4-glucosidase (lysosomal)

What is antibody-dependent cell-mediated cytotoxicity?

• Mabs bind to tumor antigen • Natural Killer (NK) cells recognize the Fc region of Mabs via Fc receptors • NK cells cause tumor cell lysis

Describe Glycosylation of EPO

•One O-linked glyc of a serine • Three N-linked glycs of asparagines