1. Identifying a Potential Biotechnology Product
6.1: Sources of Potential Products
6.2: The Use of Assays

2. Use of Plants & Animals as Sources of Biotech Products
Plasominogen activator (t-PA): dissolves blood clots
Human make it naturally in small quantities
t-PA gene is cloned in CHO cells in large quantities
Antibiotics: inhibit growth of bacteria
Produced in bacteria and fungi

3. Modeling the Research & Development of a Potential Product: Amylase
Amylase: enzyme produced by several organisms (in saliva of humans)
Breaks down starch into glucose

4. Estimating Market Size
Large market b/c industries need to easily & economically break down starch into glucose
Textile Industry: use it to remove excess starch from paper products
Beverage Co.: require large amounts of sugar & high-fructose corn syrup made by converting glucose to fructose

5. Why such a need for sugar?
Sugar was harvested from sugar cane or sugar beet
Loss of land to development = less sugar
Amylase breakdown of starch to sugar is an economical alternative to traditionally harvested sugar.

6. Identifying Amylase Product Sources
Produced in salivary glands & pancreas.
Breaks down the starch in food into absorbable sugar used as cellular fuel
Bacteria & Fungi use it to break down plant molecules for food
Bacillus subtilis: lives in soil & on decaying
plants

7. How is it mass produced?
Genencor International, Inc. genetically engineered E. coli to produce recombinant alpha-amylase on a large scale.
Why E.coli ?
Reproduces quickly
Easy to purify the product from the cell culture.

8. Creating a Comprehensive Product Development Plan (CPDP)
Does the produce meet a critical need? Who will use the product?
Is the market large enough to produce sufficient sales? How many customers are there?
Does preliminary data support that the product will work? Will it do what the company claims?
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 will it be sold for?

9. How will scientists now when they have an amylase product?
Assay
Test used to determine when they have amylase, how much, & how active it is.

10. Types of Assays Activity Assay Concentration Assay
Show compound is present and is functioning as expected.
Ex. Amylase: how much starch is broken down or how much sugar is produced
Ex. Starch/Iodine Test
Concentration Assay
Test to show the amount of molecule present in a solution
Ex. Spectrophotometer
Ex. Bradford protein reagent: shows color change

11. ELISA Enzyme-Linked immunosorbent Assay
Determines the presence and concentration of a specific protein utilizing antibody-antigen specificity.
Antibodies recognize & bind to only certain antigens which causes a color change.
Widely used in biotech labs

12. Other Assays Potency Toxicology Stability
Determine how the dosage of a drug affects its activity & how long it stays in the body
Toxicology
Show what quantities of the drug are toxic to cells, tissues, and model organisms.
Stability
At what temp, humidity, & light level should a product be stored? In what form?: liquid, powder, ect.

13. 6.1 Review Questions
Why are antibiotics important biotechnology products?
Before the widespread use of antibiotics in the 1940’s many people died from common diseases that we now treat fairly easy such as, strep throat, bronchitis, and pneumonia

14. What is the function of the enzyme amylase?
Break down the large molecule starch into smaller molecules—glucose.

15. 3. Why might a company be interested in producing amylase as a product?
Since amylase may be used to produce sugar, any industry needing large quantities of sugar would be interested. It’s easy to produce and cheap!

16. 4. Summarize the criteria that a potential product must meet in a CPDP review.
See slide.

17. 6.2 Review Questions
What kind of assay would use Bradford reagent in the test?
Protein concentration. It’s a nonspecific protein indictor and will show the presence of any protein in solution. A technician can determine the concentration of an unknown sample by comparing the unknown to known solutions

18. 2. For what purpose would a technician use an ELISA?
Determine the presence & concentration of a specific protein utilizing antibody-antigen specificity.

19. 3. What does a stability assay measure?
Show the shelf life of a product. At what temperature, humidity, and light level should the product be stored. In what form should it be stored: liquid, powder, freeze-dried, capsules, etc.

20. 4. In a large company, which department would have several employees developing and conducting assays?
An entire department. Some companies have Assay Services and Quality Control Departments that specialize in testing company products.