NOCTI: Biotechnology

What is biotechnology?

The manipulation of organisms of their parts to create products or services that benefit humankind

What are the different fields that biotechnology offers?

1. Industrial and Environmental Biotechnology

2. Medical/Pharmaceutical Biotechnology

3. Agricultural Biotechnology

4. Diagnostic Research Biotechnology

What are examples of Industrial and Environmental Biotechnology?

-fermented foods and beverages

-genetically engineered proteins for industry

-DNA identification/fingerpainting of endangered species

-biocatalysts

-biopolymers

-biosensors, bioterrorism, and biodefense

-bioremediation

-biofuels

-3D Bioprinting

What are examples of Medical/Pharmaceutical Biotechnology?

-medicines from plants, animals, fungi

-medicines from genetically engineered cells

-monoclonal and polyclonal antibodies

-vaccine and gene therapy

-prosthetics, artificial, or engineered organs and tissues

-designer drugs and antibodies

-genetic testing for human disease/disorders

What are examples of Agricultural Biotechnology?

-breeding of livestock and plant crops

-aquaculture and marine biotechnology

-horticultural products

-asexual plant propagation and plant tissue culture

-transgenic plants and animals

-production of plant fibers

-pharmaceuticals in genetically engineered plant crops

What are examples of Diagnostic Research Biotechnology?

-DNA and protein synthesis

-DNA and protein sequencing, genomics, proteomics

-genetic testing and screening

-DNA identification and DNA fingerprinting, forensics

-bioinformatics, microarrays, POLYMERASE CHAIN REACTION (PCR)

-RNAi, siRNA, miRNA

-ELISA, Western Blots, protein identification, purification

-nanotechnology

-CRISPR/Cas System

-Synthetic Biology

What can you do in biotechnology?

1. Services

2. Operations

3. Commercial Operations

4. Research and Development (R & D)

Examples of Service Careers in Biotech

-management consulting

-venture capital & banking

-law

-recruiting

Examples of Operations Careers in Biotech

-Bio IT

-Quality

-Operations & Manufacturing

Examples of Commercial Operations Careers in Biotech

-Business development

-Corporate communications

-Product

-Sales

-Marketing

-Medical affairs

-Regulatory affairs

Examples of R & D careers in Biotech

-Discovery Research

-Preclinical Research

-Bio/Pharmaceutical Product Development

-Project Management

-Clinical Development

-Regulatory Affairs

What does the lifecycle of product development look like?

1. Drug R&D

Seed Round (A/B)

A. Target ID & Validation (1-2 Yrs):

-link disease and target

-Biomarkers

-High thought out screening

-Rational design

-In silicon screening

B. Hit Generation (1-2 Yrs):

-Hits Confirmation

-Potency & Cytotoxicity

-Prelim animal efficacy

-Initial SAR

Series A (C)

C. Lead Gen and Optimization (1-2 Yrs):

-Potency Studies

-Selectivity Studies

-PK/ADME-Tox properties

-SAR pharmacophore modeling

---

Series B (D)

D. Pre-Clinical Animal Studies (1 Yr):

-Pharmacoligical profile

-Administration route

-Drug interactions

---

2. Clinical Trials

Out Licencing or Series C (A)

A. Phase 1 Safety (1-2 Yrs): 20-100 People

B. Phase 2 Efficacy Safety (1-2 Yrs): 100-300 People

C. Phase 3 Efficacy Safety (1-2 Yrs): 1,000-3,000 People

---

3. FDA Review and Approval (1-2 Yrs)

What does R&D entail in product development?

Seed Round (A/B)

A. Target ID & Validation (1-2 Yrs):

-link disease and target

-Biomarkers

-High thought out screening

-Rational design

-In silicon screening

B. Hit Generation (1-2 Yrs):

-Hits Confirmation

-Potency & Cytotoxicity

-Prelim animal efficacy

-Initial SAR

Series A (C)

C. Lead Gen and Optimization (1-2 Yrs):

-Potency Studies

-Selectivity Studies

-PK/ADME-Tox properties

-SAR pharmacophore modeling

In summary, finding the drug target, making the actual drug, identifying and purifying the small molecule, scaling it up, and doing lab-based research on nonliving organisms.

What is the purpose of pre-clinical trials?

D. Pre-Clinical Animal Studies (1 Yr):

-Pharmacological profile

-Administration route

-Drug interactions

The main goals of preclinical studies are to determine a starting, safe dose for first-in-human study and assess potential toxicity of the product, which typically include new medical devices, prescription drugs, and diagnostics. As we work with animals in this case, we have model organisms to see the impact of the products.

What happens during Target ID and Validation?

Target ID & Validation (1-2 Yrs):

-link disease and target

-Biomarkers

-High thought out screening

-Rational design

-In silicon screening

How long does each process in product development take on average?

1-2 Yrs

What happens during the hit generation?

Hit Generation (1-2 Yrs):

-Hits Confirmation

-Potency & Cytotoxicity

-Prelim animal efficacy

-Initial SAR

This is where a small molecule that interacts with a biological target is evaluated and undergoes limited optimization to identify promising lead compounds. Then, its potency and cytotoxicity are analyzed. This sometimes happens on animals

What is the Seed Round?

This is the financing round that raises initial capital to start a business. In biotech, this consists of these two steps:

1. Target ID and Validation

2. Hit Generation

This is because this is the time where a compound and problem are identified and tested to see if there is an idea that can be built upon.

What two steps make up the Seed Round?

1. Target ID and Validation

2. Hit Generation

What happens during Lead Gen and Optimization?

Lead Gen and Optimization (1-2 Yrs):

-Potency Studies

-Selectivity Studies

-PK/ADME-Tox properties

-SAR pharmacophore modeling

What does Series A consist of?

Lead Gen and Optimization (1-2 Yrs):

-Potency Studies

-Selectivity Studies

-PK/ADME-Tox properties

-SAR pharmacophore modeling

What does Series B consist of?

D. Pre-Clinical Animal Studies (1 Yr):

-Pharmacoligical profile

-Administration route

-Drug interactions

What happens during Pre-Clinical Animal Studies?

-Pharmacological profile

-Administration route

-Drug interactions

What are you checking for in each phase of the clinical trials?

Phase 1: Checking for Safety

Phase 2: Checking for Efficacy

Phase 3: Confirm Findings in Large Patient Population

Phase 4: Testing Long-Term Safety in Diverse Patient Populations

What happens during Phase 1 of Clinical Testing?

Phase 1: Checking for Safety

You start with HEALTHY PEOPLE

testing a new drug on a small group of healthy individuals. to find out what a safe dosage is, if there are any side effects and how the body reacts to drug = SIDE EFFECTS

20-100 People

1-2 Years

What do you start with in Phase 1?

healthy people

How large of a group can you test on in Phase 1?

20-100 people

What happens during Phase 2 of clinical trials?

Phase 2: Checking for Efficacy

Proof of concept, finding beneficial/undesirable effects

100-300 people

Diagnosed patients

1-2 years

Testing for dosage

What kind of people do you work with in Phase 2 of clinical trials?

Diagnosed patients

How many people can you work with in Phase 2 of clinical trials?

100-300 people

What kind of effects are you looking for in Phase 2 of Clinical trials?

Proof of concept, finding beneficial/undesirable effects

testing for dosage

How long is Phase 1 of CT?

1-2 years

How long is Phase 2 of CT?

1-2 years

What happens during Phase 3 of clinical trials?

Phase 3: Confirm Findings in Large Patient Population

-comparative efficiency: testing how it compares to existing therapies and drugs

-looking for statistical significance: is it worthwhile?

-1,000 to 3,000 people

-2-3 years

What are you testing in Phase 3 of clinical trials in comparison to Phase 2?

Phase 3 comparative efficiency: testing how it compares to existing therapies and drugs while Phase 2 was focused on if the effects were generalizable

What can be realized in Phase of CT?

statistical significance, is it worthwhile, of a product

How many people are tested on in Phase 3 of CT

1000-3000 people

How long does Phase 3 of CT take?

2-3 years

What happens during Phase 4 of Clinical Trials?

Testing Long-Term Safety in Diverse Patient Populations

FDA Review and Approval - Long Term Side effects

What happens after the FDA New Drug Application and Approval?

long term effects are studied for

What is an independent variable?

factor in an experiment that a scientist purposely changes

What is a dependent variable?

The dependent variable is the variable that is measured

What is a control group?

The group that does not receive the experimental treatment in an experiment.

What is an experimental group?

the group in an experiment that receives the variable being tested

How are experimental controls different than the control group?

Experimental controls mean making sure that the only thing being changed is the independent variable.

Control group is the group that does not receive the experimental treatment

Experimental Controls

Experimental controls mean making sure that the only thing being changed is the independent variable to prevent skewed results/correlations.

What is the equation for molarity?

M = mol/L

Equations for Concentration

Concentration to grams : (g/mL) * (volume) = grams

If you are given a %... change to a decimal first

Decimal is then the concentration (g/mL)

How to change from the concentration to grams?

(g/mL) * (volume) = grams

What happens if you are given a concentration as a %?

If you are given a %... change to a decimal first

Decimal is then the concentration (g/mL)

If you need to find the mass, multiple by volume

What equations do you use for buffer calculations?

(g/ml) * (mL) = g

What is the equation used for dilutions?

C1V1 = C2V2

What makes a good experiment?

Randomized, controlled

Random assignment into groups

Experimental controls → making sure that the only thing being changed is the independent variable

Independent variable → what you're changing

Dependent variable → the response to the independent variable

What does it mean for an experiment to be randomized?

Random assignment into groups

What does it mean for an experiment to be controlled?

Experimental controls → making sure that the only thing being changed is the independent variable

Independent variable → what you're changing

Dependent variable → the response to the independent variable

How much solute is needed to make 550 mL of a 2.7 g/L gelatin solution?

0.550 L * 2.7 g/mL = 1.485 g

How much 95% Ethanol do you need to make a 600mL solution of a 70% ethanol solution?

C1V1 = C2V2

95(x) = 70(600)

x=442.105 mL

Making Solutions of Differing Mass/Volume Concentrations

Concentration desired (g/mL) * Total Volume Desired (mL) = Mass of solute in the total volume desired

How to prepare a Mass/Volume Solution

1) Measured mass of solute

2) Measured volume of solvent to the desired volume

3) Slowly add solute to solvent, allowing time for the solvent to absorb

CuSO4 Solution To be Bade

5.0 mL of 300 mg/mL

4.5 mL of 150 mg/mL

4.0 mL of 75 mg/mL

3.5 mL of 37.5 mg/mL

3.0 mL of 18.75 mg/mL

Making Solutions of Differing % Mass/Volume Concentrations

Step No. 1:

Convert the % to a decimal:

% percent value = decimal value of the g/mL

Step No. 2:

decimal (g/mL) of the % concentration * volume (mL) = grams of solute to be measured. Add solvent to solute until the desired volume is reached

Prepare 5 mL each of 5%, 2,5%, 1,25% and 0.625% CuSO4 pentahydrate solution

40 mL of 6.5 mg/mL CuSO4 pentahydrate solution

200 mL of 8% NaCl solution

0.75 L of 5% dextrose solution

10mL of 1.25% NaOH solution

150 mL of 0.5% NaCl solution

Making Solutions of Differing Molarity Concentrations

Volume Wanted (L) Molarity Desired (mol/L) Molecular Weight of the solute (g/mol) = grams of solute to be dissolved in solvent to the final desired volume

550 mL of 9.5 mg/mL NaOH solution

150 mL of 2% CuSo4 * 5H20 solution

3.0 L of 0.025% dextrose solution

125 mL of 10M NaOH solution

75 mL of 0.1M NaCL solution

25 mL of 10 mM NaCl solution

Making Dilutions of Concentrated Solutions

CV=CV

C1 = {} of starting solution

V1 = v to use of stock solution in the diluted sample

C2 = desired {} of starting solution

V2: desired volume of the diluted sample

Making a 1:2 Serial Dilution

divide initial concentration by 2

50 mL of 15 mg/mL NaOH solution from 100 mg/mL NaOH

10 mL of 0.5 CuSO4 5H20 solution from 10 M CuSO4 5H20

2 L of 5 mg/mL gelatin solution from 1 g/mL gelatin

950 mL of 1X CuSO4 5 H20 solution from 25X CuSO4 5H20

5 L of 0.2M dextrose solution from 5 M dextrose solution

100 mL of 2.5 NaOH from 50x NaOH

50 mL of 5 mM NaCL from 1 M NaCl

Given the following buffer solute concentrations, how would you prepare 100mL of this buffer?

1.9 mM NaH2PO4 ᐧ H2O

137.99 g/mol

8.1 mM Na2HPO4

141.98 g/mol

0.5 M KCl

74.55 g/mol

pH = 7.35

What does a standard curve help us do?

determine unknown concentration using line of best fit

What does an absorbance and concentration graph look like?

X axis = concentration (g/L)

Y axis = Absorption (au)

Plot all the points

Find the Linear Regression Equation y=ax + b

Find the R^2 value

How to find the R^2

Before you do your standard curve graph, click 2nd 0

Click x^-1 to get to CATALOG

Scroll down to DiagnosticOn and turn that on with enter

Put in your L1/L2 in Stat and FInd the Linear Regression Equation

Input vars for the Y1 Function

Correlation Coefficient (r)

A single number, ranging from -1.0 to 1.0, that indicates the strength and direction of an association between two variables.

coefficient of determination (r^2)

The fraction of the variation in the values of y that is accounted for by the least-squares regression line of y on x.

if r < 0 then

there is a negative linear relationship between the 2 variables (slope of the regression equation is negative)

If r <0 then

there is a positive linear relationship between the 2 variables (slope of the regression equation is positive)

The sign of r indicates?

The magnitude of r indicates?

The sign of r indicates the direction of linear relationship between the 2 variables

The magnitude of r indicates the strength of the linear relationship between the 2 variables

If r=0?

If abs value of r=1?

If r=1 or -1?

If r=0, then there is no linear relationship between the 2 variables

If abs value of r=1, then there is a very strong linear relationship between the 2 variables

If r=1 or -1, hen there is an exact linear relationship between the 2 variables

0 < r^2 < 1

The coefficient of determination or R^2 measures how close the data values are fitted to the regression line. It measures the proportion of the variance in the dependent variable (output) that is predictable from the independent variable (input)

when is r^2 significant

In other fields, the standards for a good R-Squared reading can be much higher, such as 0.9 or above. In finance, an R-Squared above 0.7 would generally be seen as showing a high level of correlation, whereas a measure below 0.4 would show a low correlation.

What is the formula/rules for writing a title for the Standard Curve graphs?

An Analysis of Absorbance (au) at 590 nm of various concentrations of Copper (ll) Sulfate Pentahydrate (g/mL)

Comparison of different solution preparation techniques with differing concentrations of CuSO4*5HO vs. Absorbance at 590 nm (Au)

Given that the 3g/L solution has an absorption of 0.6au - if you performed a 1:4 dilution on it, what would be the expected absorbance of the solution?

1:4 dilution = divide initial concentration by 4

If you know the absorbance, use regression line to give what the suspected concentration would be

3/4 g/L

0.6/4 = 0.15 au

What should you have in your graph?

X Y labels

data points

legend

title

least squares regression line (LSRL)

How would you determine the [ ] of an unkn soln?

Through extrapolation, using the LSRL formula you got from your calculator and plugging a variable in

How does a spectrophotometer work?

Measures light absorption or amount of chemicals in a solution.

Uses a light beam which passes through the sample, and each compound in the solution absorbs or transmits light over a certain wavelength