Antibodies and Gene Therapy Flashcards by Jon Beckloff

Monoclonal Abs mAbs

Polyclonal Abs

3 reasons not why

What do you need though?

What is the problem?

Even if a B cell produces what you want?

What do we need?

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Middle Ear catheter

Hows it work?

Is it invasive?

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Examples of ELISA tests

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Compatibility of Protein drugs with other delivery devices (PUMPS)

Reports of adsorption of protein to tubing in IV infusion: 3 things Drugs that were effected
What could the compatibility affect? 2 things
Considerations should be followed just like?

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Protein Composition

What type of chain and what is it made up out of?

How many naturally occuring types?

What are 4 ways their chemical makeup varies?

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Comparison of Genome Editing Tools

3 on the slide

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Genome editing

How to cut DNA Precisely at a desired location

One of this is pretty common and uses a metal

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Examples of Variables to Consider during IV admin of Protein Therapeutic DRugs

What I just said but examples

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Example 1 continued

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Protein folding

Driving force based on what sequence?

4 points? Vs. What other point?

Collapse?
What interactions?
What bonding?
This list is not?

Vs.

Protein and water? Entropy?

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Humoral Immunity and Antibody

Critical for protection against? Or for?

How to most vaccines elicite protection?

Antibodies in Human healthcare

How many approved Ab drus?

How many in clinical trials?

How many approved vaccines?

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Antibody half life extension

This is talking about how protection is transfered to a baby.

IgG binds to?

How? what is the mechanism?

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Other otic diseases?

5 different ones

Which one can be given steroids and antioxidance prophylaxtically

Sudden Sensoroneural Hearing loss

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Goals for Formulation

The protein needs to be stable how do you do that? 2

What factors do you need to consider with parenteral forms? 4 things

Why are delivery devices important? and why? what type of things can happen? 2 reasons

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Catheters- Cochlear Implant

What have they developed in recent years?

To prevent rejection of the wires the coat them with glucocorticoids that slowly release to reduce inflammation and build up of scar tissue

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Generating human Abs

3 ways to accomplish this

Humanization- Loop sequence from mouse tranplanted to human Ab and doesnt lose affinity- Less immunogenic, Really want to make completely human Abs
Human Abs from transgenic (humanized) mice- These have a human immune system, But sometimes they cant make Abs with high enough affinity for the disease
Generation of Human Abs from Combinatorial libraries in vitro 2 kinds can be make Phage display technique, Isolation of spleen cells PCR amp, Ab genes are then exprssed in bacteria the 2 libraries- From healthy unimmunized people, Biased libraries made from immunized individuals There are also synthetic libraries

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Model of Multi state process of Non-Native Protein aggregation

What is the best and most effective form of the protein?

Because of this what do you want to prevent?

The goal is to keep the protein in as mich of the monomer form as possible or to have it readily be reversed back into the monomer so the drug can still be active

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Anti Tumor Vaccines

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Long t1/2 application Agglutination test

Fancy word for?

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Protein Folding

We say that it cannot be a? What paradox?

What about the timescale of protein folding?

Therefore? 3 points

What type of folding and what type of intermediates?
What do solvent interactions do?
What type of proteins in vivo

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Formulation stresses affecting protein aggregation

Role of Formulation variables 3 things

Different Classes of?
Protein?
Multi-use?

Role of interfaces 3 things with examples

Liquid to
Liquid to
Liquid to

Role of Processing 3 things with examples

Operations?
Keeping something still?
When you are giving it

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Ear anatomy the middle and inner ear

and zoomed in on the Cochlea

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Potential Gene and Cell Therapy Targets

What are the 4 types of targets?

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Microwick

What does it do?

And how does it work?

Is it invasive?

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Drug Labels: Selected sections that comment on Storage and Stability * Simponi must be? * Do not use beyond? * Do not? * Do not? * Simponi is provided in? * Simponi doesnt contain? * When should you not use the solution?

TALENs?

Transcription Activator-like effector nuclease

Hybridoma tech 7

Reversible self association of mAbs at high protein concentration What are the interactings between the mAb monomers that allow them to be reversible? 3 one is obvious

Monoclonal Abs uses in biotech and medicine 12 things? Drug examples 2

B cell Hybridomas Secrete monoclonal Antibodies Polyclonal B cell problem Fuse B cells?

Bad picture of middle ear

Mechanism of Protein Stabilization by Certain Solvent additives When sucrose is preferntially excluded from the surface of proteins what happens? Which type of protein has more surface area? According to Le Chats principles? What do proteins do to overcome thermodynamically unfavorable conditions?

Modeling Inner ear Drug delivery Concentration gradients and graphs

Bottom is distance Stating with cochlea

RWM Structure * Outer * What type of cells? * Rich in what two things? * Continuous with? * Junctions? * Middle * When is elastic density highest? * What 3 things are in this? * Inner * What type of layer epithelial shape with what type of extensions?

Transposase Mediated Gene Delivery

Protein stabilization using Excipients Chemical class 7 MOA 6

Examples of post-translational modifications and degradation of proteind in vivo 7 things

Examples of commercial products high protein concentration SUBQ delivery 6

Protein Degradation pathways In Vitro Chemical Pathways 8 Just know the differences between the two Physical Pathways 4 and 4 subs in last one

Sterile prep of protein drugs and IV Admin Sterile processing during manufacturing of protein drugs usually involves? Sterile handling may involve? 2 things Stability within?

Immuno PCR What does it do what is it able to detect? Combines what 2 tests? This provides extremely?

Whats the point to developing High Concentration mAb formulations? Sub Q admin typically requires? 1. mAb range? 2. How is the SubQ injection limited? 3. How is at home use facilitated? What are 5 pharmaceutical challenges involved? 1. If it will disolve 2. If it is too? 3. Realistic? 4. Stability in? 5. _____ related to 2nd point

ELISA

Compatibility of New formulations designed for at home use 2 brand names One is Lyapholized and one is a solution What are some issues that at home use can cause for stability

Clinical trade off between Control of Drug level and Risk picture chart thing 7 different modifications

ELISA Variants

Hybridoma Technology Whar are the total of 7 things things you need?

1. Mouse spleen that is immnized with what you want 2. Use agents to facilitate fusion of membranes 1. Low success rate here, Myeloma cells dont have HGPRT, these myelomas cannot be able to synthesize Ab molecules are there own 3. Unfused cells cant grow because they dont have HGPRT, unfused normal spleen gant grow forever because of life span, Hybridoma can grow because it has the HGPRT needed for the myeloma cell to survive adn the meyloma partner doesnt die 4. Find the ones producing Ab 5. Isolate single cells from each ab(+) culture and subculture those for specificty 6. Each (+) subculture is now a bunch of clones and they are monoclonal 7. Now scale up

IV delivery and protein stability Setup varies in 4 ways 1. ___ and ___ rate 2. Size of 3. \_\_ 4. \_\_ IV bags 2 Different? and Different types of? Admin kits 2 Different types of? and Different?

Hearing Picture of cochlear frequency

People can damage their hearing at different sites and this is specific to these regions

Protein Stability Which state is more stable? Unfolded or Folded? Numerically how much more stable? 4 points pertaining to this? 1. How many Hydrogen bonds? 2. What about small proteins? 3. What is only marginally stable? 4. Unfolding? Protein stability refers to?

More cochlear anatomy What is inside the cochlea? Why is this important for drug delivery?

Introduction Recombinant proteins made from different cell lines leading to? AS in what are the 3 types of organisms used for protein things 3 cell types used Proteins are Sensitive what 4 things are they sensitive to? What makes them not work Formulation Turning molecules into medicine 4 things 1. What do you do for clinical admin? 2. What do you need to figure out when the pt is taking it? 3. How lng can it last? 4. Friendly to who?

Transposase Mediated Gene Delivery

Developing a Stabilizing formulation is Essential Start with formulation considerations 4 things in the middle Then you end with?

Role of environmental stresses? Role of solution conditions? Effect of environment stresses and solution conditions on: what 4 things?

Cochlear crosssection

Monoclonal antibodies picture What is the difference between Small Drugs and Large biologics in terms of how they are made up

The antibody universe Humans have the potential to make how many antibodies? Are antibodies common? Antibody genese in newly formed? Formed where? They are assembles from combinatorial stitching of 3? and 2? Newly Form B cells Naive Bcells How many in the blood? Encode how many antibodies? Bind molecules with? Are they selective? Cells displaying antibodies that bind to "self"? When does a Cell become inert?

Passive Serum Transfer Antibody serum or purified immunoglobulin transferred to another to? Horse serum is stilled used for? How can human blood products be used? These are used to provide what type of immunity? and to what type of peopl? Example? IVIG is manufactured from?

Evolution of Antibody Technology

Dynamic Behavior of Immunoglabulins What region connects Immunoglubulin fragments? 2 parts of this? What is between V and C domains? 2 types of motions Sidechain?

Applications of CRSPR tech

What does a long half life mean for the drug? Relieves pressure for development in 6 ways What does this do overall?

Ven Diagram Preformulation Characterization: Determine Degradation Pathways and Physiochemical Stability Profile 4 parts The end thing in the middle is?

Comonly Prescribed Otics Antibiotics? 3 points Analgesics 1 Something that losens earwax? Steroids? Antifungals?

CRISPR

Cluster Regularly Interspaced Short Palindromic Repeats

Abs structure and recognition? What does IgG consist of? WHat are the variable regions? Treating the Ab with what can cleave it into what 2 parts?

Generating human Abs 2nd way out of three

Substances that cross the RWM Some are weird whY?

Ferritin, Albumin, latex spheres very big its weird they can get in

Monoclonal antibody structure 2 portions? Important physical characteristics 5 1. How heavy? 2. How many AAs? 3. Domain? 4. Sugar? 5. Type of molecule?

Modeling inner ear drug delivery LADME? RWM is like? How does delivery work?

Essentially a very small hole in a narrowing hallway

Infusion hypersensitivity Rituximab example What type of Ab is this? \_\_\_/\_\_\_ anti? What were the 2 reactions involved? two main causes? 1. Rapid targeting of cells for destruction causes rapid immune response 2. Immune response to protect therapeutic

Elisa Results

Why do we care about RSA od therapeutics? Big ass chart

Examples of Protein drug Heterogeneity? 3 Question is what makes the proteins vary non hemogenous

Protein Physical Stability and Aggregation Two general categories: Two roles

Non- Ionic surfactant minimizing? What is an example of this?

What is formulation development for protein drugs? 1. Drug at the appropriate? 2. Excipients to ensure? 3. Primary? 4. For admin? 5. What type of manufacturing process? 6. Develop what methods? 7. Define what conditions?

Protein stabilization using excipients divided into what two things?

Chemical Class and MOA

Stability challenges to Develop [high] liquid mAbs in prefilled syringes 2 things 1. Proteins in what type of formulation? 1. These are usually? 2. Solubility limite? mg/ml 3. 3 issues 2. Compatability of ___ protein formulation with? 1. Componenets of? 2. Silicone and Tungten 3. What type od device?

The RWM as a gateway to the inner ear * How long has the RWM been observed? * Why is this an attractive drug target?

TIL: Stand for? What do you need to isolate? CAR: Stans for? These are engineered? What do they express? What happens when they bind? In vitro reprogramming of T cells to recognize tumore cells: Isolated T cells are incubated with? These have been fed wiht? What does this stimulate and activate? These are then expanded to?

Transposase Mediated Gene Delivery What are the two technologies? how do they work?

Example 1 of mAb RSA

T cell therapies for cancer Tx

What are the 4 protein structure types? What are those made up of?

How can mAbs eliminate tumors 3 ways

Advantages of Immunoassays? 5 things 1. How small of detection? Meaning it is? 2. How can it be done? can you do it again? 3. What do you need? 4. What type of data? 5. What about automation?

Genetics and Antibody production What are the constant and variable regions?

Antigenicity? Immunogenicity? All molecules that have property of Immunogenicity also have property of? What isnt true? Hapten? What are the 4 factors that affect a substances immunogenicity?

Phage display What type od technique is this?

3rd way Human Monoclonal

Factors influencing Immunogenicity of Therapeutic Proteins Picture of a bunch of stuff

Protein Met Oxidation Protein Trp oxidation

Physical degradation pathways of protein therapeutics 5 things and these cause what 2 things? What is the solution to the problem?

Stresses Affecting Protein Physical Stability two general categories

Epidemiology of hearing loss * How common is congenital hearing loss? * Some studies suggest? * What percentage is syndromal? * Nonsyndromal what percentage is from recessive gene and what percentage dominant?

Intratympanic long acting gels how do they work?

Needle of solution in through the tympanic membrane and edge of round window once temp increases within the boudy the solution turns into a gel and sticks to the Round Window and can be absorbed due to time at active site This is a less invasive fix

Mechanism of Protein stabilization by certain solvent additives Examples of stabilizing Solvent Additives 3 Common view? Stabilization properties are not due to? 2 other things

Examples of Chemical Degradations pathwzys Protein?

Chimeric Antigen Receptors (CARS) WHat are they What is difficult about them? What reactions happen? Pros?

What does the constant region do? What does the variable region do?

Humoral Immunity: The ensemble of Ab in the serum and in secretions that confer protection against disease What are vaccine- Antibodies are elicited by? Many of which contribute to? What dont we know? What are therapeutic Abs? Discovery of? That are responsible for?

Antibodies for commercially useful dianostics What 4 properties does a detection system need? What do these mean?

WHy are murine mAbs not ideal for therapeutics? NonHuman Abs are only used in two special cases?

Factors affecting RWM permeability 3 things Membrane Inflammation What is passing through?

Picture of organ of corti? With lots of vocab What are the two types of hair cells? What is inside the hair cell? What are the 3 modulators? 3 causes of Organ of Corti Damage? 4 types of therapy?

The hair cell only have one nucleus so it makes sense that they can die quickly

Antigen what does it mean? What are the 5 ones in the slide?

Ethical Questions in Gene therapy? Are disabilities diseases? Do they need to be cured or prevented? Does searching for a cure demean the lives of individuals presently affected by disabilities? Is somatic gene therapy (treating adult cells of persons known to have the disease) more or less ethical than germline gene therapy (which is done in egg and sperm cells and prevents the trait from being passed on to further generations)? Preliminary attempts at gene therapy are exorbitantly expensive. Who will have access to these therapies? Who will pay for their use?

Antidrug Antibody detection Bridging? Where is it detected? What is the critical factor in this?

Human Humoral Immunity Mining abs from?

Ag binding Loops of? What makes the Ag Ab binding tigher? Typically therapeutic Ab binding? What are the 4 forces responsible for Ab to Ag binding? Abs that bind to proteins form a binding site that is? Abs that bind to small molecules?

Compatibility of Protein drugs during IV admin 3 things 1. Manufacturers test for compatibility so carefully review dosing and admin instructions to know: 1. What to use 2. And the need for? 2. Need to know what it looks like? 3. Need to know?

Gene therapy What is the promise? 4 problems in therapy 1. What is hard to deliver? 2. Other issue with delivery? 3. What are some effects that can occur? What has fewer of these? 4. 2 things about the engineered cells

Regulatory Guidelines ICH-M4Q USP797 Address proper reconsitution

Antibody half lfe what has recently been done? Mutations of Fc variants what does this do? 2 things

Role of conformational stability? 2 points Role of Colloidal Stability 2 points? What happens as a consequence of attractive intermolecular forces? What influences proximity?

Factors affecting RWM transit What impairs latex spheres to cross? What type of ferritin can cross and what type cant? What 4 substances can increase permeability?

Genome Editing Methods for Precise insertion or deletion of DNA fragments in Eukaryotic Cell s 3 things

Examples of Chemical Degradation Pathways 5 and examples within those

Formulation stresses affecting protein aggregation 3 roles

Role of: Formulation varriables, interfaces, and processing

From Naive B Cells to Antibodies The entire question is how do Naive B cells go from niave to ones that produce antibodies? How are bacterial molecules presented to B Naive B cells? What are activated B cells and what do they do? What does this introduce? B cells that display higher affinity for the antigen what happens? Ultimately what happens to B cells that win? What 2 things are they called?

Route of Admin for FDA approved mABs So what types of routes are most common

IV, Sub Q, other most to least

The dynamic Immune System: Continuous Decay and? How much does the human immune system weigh? How many lymphocytes? How many antibody molecules? During this lecture how many?

Protein Aggregation

Gene Therapy Definition Insertion of a ___ \_\_\_\_ into the cells of a patient thereby ____ an ____ \_\_\_\_, or conferrring?

Systemic approaches to drug delivery Flow chart

Antibodies and Gene Therapy Flashcards

(127 cards)