Transcript Identifying a Potential Biotechnology Product

Identifying a Potential
Biotechnology Product
Chapter 6
Objectives
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Describe the CPDP process.
List and describe the different assays that can be
performed on proteins.
Describe the processes that can used to isolate
molecules of interest from plant cells.
Describe how plant proteins of interest can be
mass produced by cells.
Is it worth pursing a product or not?
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Biotechnology products can come from a variety
of sources.
Decision needs to be made as to whether it is
worth pursuing the development and sale of a
product.
Many companies use a CPDP strategy.
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Comprehensive Product Development Plan
Is it worth pursing a product or not?
CPDP
1.
2.
3.
4.
5.
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Does the product meet a critical need? Who will use the
product?
Is the market large enough to produce sufficient sales? How
many customers are there?
Does the preliminary data support that the product will work?
Will it do what the company claims?
Can patent protection be secured? Can the company prevent
other companies from producing it?
Can the company make a profit on the product? How much
will it cost to make? How much can it be sold for?
I.e. common cold vaccine
Amylase
Verifying what you have.
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A majority of the biotechnology products
produced are proteins.
Must be able to determine:
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Is product present?
What is the overall activity of the product?
Concentration of product.
Difficulty in working with proteins is that they
are colorless and nonvisible to the naked eye.
Assays
(Tests)
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1. Activity assays
2. Concentration assays
Bradford protein reagent or bicinchoninic acid
(BCA)
 Enzyme-linked immunosorbent assay (ELISA)
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Verifying what you have.
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Generally assays,
meaning test, are done
when working with
proteins.
Activity assay
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Indicate presence
Indicate whether
protein is working or
not
Lighter color…..more active amylase
Negative
Verifying what you have.
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Concentration assay
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Amount/unit volume of a protein
By eye or spectrophotometer
Bradford protein reagent or
bicinchoninic acid (BCA)
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Nonspecific in its action
ELISA
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0.01, 0.1,
1mg/mL
Decreasing amount of
protein…..indicated by less blue color in
tube
Enzyme-linked immunosorbent assay
Very specific
Unitize antigen-antibody specifity
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An enzyme attached to an antibody binds
to a specific protein (antigen)
Causes a color change that can be meas.
When specific substrate is added
The darker the color…..higher conc.
Of protein
Verifying what you have.
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Dealing with pharmaceuticals
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The stringency of verifying what you have, that it is safe, and that it
works are much higher.
In addition to activity and conc.
PK assay
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PD assay
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Pharmacodynamic (length of activity)
Additional assays developed and carried out to show potency,
toxicology, and stability of drug.
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Pharmacokinetic (amount of activity of protein in humans and other test
organisms)
Potency: how dosage of a drug affects its activity and how long stays in body
Toxicology: what quantities of drug toxic to cells, tissues, and model
organism
Stability: shelf life, and storage conditions
Assay Services and Quality Control Department.
Focus on Plant Proteins as Possible
Products
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Extracting Protein Molecules from Cells
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Most molecules of interest are found within the
cell.
Problems encountered when attempting to isolate
RNA, DNA, or proteins from plant cells.
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Plant material is dense and can be quiet woody.
Cells that make up plants have cell walls that are very
thick.
Plant material broken into smaller components to
increase surface area for solvents to work on.
Focus on Plant Proteins as Possible
Products
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Removing or Weakening Cell Walls
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Enzymatic removal of cell wall
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Major components of cell wall are cellulose
and pectin.
Cellulase and pectinase enzymes degrade
cellulose and pectin.
Once cell wall removed protoplast forms.
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Plant cell with only a plasma (cell) membrane.
Homogenization of plant cells
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Crushing action that results in release of
intracellular contents.
I.e. Tissuemizer
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Works like blender
Grinds tissues without causing foam
Focus on Plant Proteins as Possible
Products
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Removing or Weakening Cell Walls
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Freeze facture
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Cells flash frozen with liquid nitrogen or dry ice
Crushed using a mortar and pestle
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Results in maceration of cells
Isolating Proteins
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Cell contents placed in a neutral solution and proteins
extracted.
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i.e. potassium phosphate buffer
Other extraction methods that can be used are acetone or
ammonium sulfate.
Once protein(s) isolated assays are generally run.
Focus on Plant Proteins as Possible
Products
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Once a particular protein has been identified a
possible way to make more of it is to produce
rDNA.
Cells can be transformed with the rDNA and begin
the process of making the protein.
Homework
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Review questions page 168
 Questions
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1, 2, 3, and 4
Think like biotech page 180
 Questions
3, 5, 6, 7, and 9
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