Identifying a Potential Identifying a Potential Biotechnology Biotechnology

ProductProduct

6.1: Sources of Potential 6.1: Sources of Potential ProductsProducts

6.2: The Use of Assays6.2: The Use of Assays

Use of Plants & Animals as Use of Plants & Animals as Sources of Biotech ProductsSources of Biotech Products

Plasominogen activator (t-PA)Plasominogen activator (t-PA): : dissolves blood clotsdissolves blood clots– Human make it naturally in small quantitiesHuman make it naturally in small quantities– t-PA gene is cloned in CHO cells in large t-PA gene is cloned in CHO cells in large

quantitiesquantities

Antibiotics: inhibit growth of bacteria– Produced in bacteria and fungi

Modeling the Research & Modeling the Research & Development of a Potential Development of a Potential

Product: AmylaseProduct: Amylase Amylase: enzyme produced by Amylase: enzyme produced by

several organisms (in saliva of several organisms (in saliva of humans)humans)– Breaks down starch into glucoseBreaks down starch into glucose

Estimating Market SizeEstimating Market Size

Large market b/c industries need to Large market b/c industries need to easily & economically break down easily & economically break down starch into glucosestarch into glucose– Textile IndustryTextile Industry: use it to remove : use it to remove

excess starch from paper productsexcess starch from paper products– Beverage Co.Beverage Co.: require large amounts : require large amounts

of sugar & high-fructose corn syrup of sugar & high-fructose corn syrup made by converting glucose to fructosemade by converting glucose to fructose

Why such a need for sugar?Why such a need for sugar? Sugar was harvested from sugar Sugar was harvested from sugar

cane or sugar beetcane or sugar beet– Loss of land to development = less Loss of land to development = less

sugarsugar

Amylase breakdown of starch to Amylase breakdown of starch to sugar is an economical alternative to sugar is an economical alternative to traditionally harvested sugar.traditionally harvested sugar.

Identifying Amylase Product Identifying Amylase Product SourcesSources

Produced in salivary glands & Produced in salivary glands & pancreas.pancreas.– Breaks down the starch in food into Breaks down the starch in food into

absorbable sugar used as cellular fuelabsorbable sugar used as cellular fuel Bacteria & Fungi use it to break down Bacteria & Fungi use it to break down

plant molecules for foodplant molecules for food– Bacillus subtilisBacillus subtilis: lives in soil & on decaying

plants

How is it mass produced?How is it mass produced?

Genencor International, Inc. Genencor International, Inc. genetically engineered genetically engineered E. coliE. coli to to produce recombinant alpha-amylase produce recombinant alpha-amylase on a large scale.on a large scale.– Why Why E.coliE.coli ? ?

Reproduces quicklyReproduces quickly Easy to purify the product from the cell Easy to purify the product from the cell

culture.culture.

Creating a Comprehensive Creating a Comprehensive Product Development Plan Product Development Plan

(CPDP)(CPDP)1.1. Does the produce meet a critical need? Who Does the produce meet a critical need? Who

will use the product?will use the product?2.2. Is the market large enough to produce sufficient Is the market large enough to produce sufficient

sales? How many customers are there?sales? How many customers are there?3.3. Does preliminary data support that the product Does preliminary data support that the product

will work? Will it do what the company claims?will work? Will it do what the company claims?4.4. Can the company prevent other companies Can the company prevent other companies

from producing it?from producing it?5.5. Can the company make a profit on the product? Can the company make a profit on the product?

How much will it cost to make? How much will How much will it cost to make? How much will it be sold for?it be sold for?

How will scientists now when How will scientists now when they have an amylase product?they have an amylase product? AssayAssay

– Test used to determine when they have Test used to determine when they have amylase, how much, & how active it is.amylase, how much, & how active it is.

Types of AssaysTypes of Assays

Activity AssayActivity Assay– Show compound is present and is functioning Show compound is present and is functioning

as expected.as expected. Ex. Amylase: how much starch is broken down or how Ex. Amylase: how much starch is broken down or how

much sugar is producedmuch sugar is produced Ex. Starch/Iodine TestEx. Starch/Iodine Test

Concentration AssayConcentration Assay– Test to show the amount of molecule present Test to show the amount of molecule present

in a solutionin a solution Ex. SpectrophotometerEx. Spectrophotometer Ex. Bradford protein reagent: shows color changeEx. Bradford protein reagent: shows color change

ELISAELISA

Enzyme-Linked immunosorbent Enzyme-Linked immunosorbent AssayAssay– Determines the presence and Determines the presence and

concentration of a specific protein concentration of a specific protein utilizing antibody-antigen specificity.utilizing antibody-antigen specificity.

– Antibodies recognize & bind to only Antibodies recognize & bind to only certain antigens which causes a color certain antigens which causes a color change.change.

– Widely used in biotech labsWidely used in biotech labs

Other AssaysOther Assays

PotencyPotency– Determine how the dosage of a drug affects its Determine how the dosage of a drug affects its

activity & how long it stays in the bodyactivity & how long it stays in the body ToxicologyToxicology

– Show what quantities of the drug are toxic to Show what quantities of the drug are toxic to cells, tissues, and model organisms.cells, tissues, and model organisms.

StabilityStability– At what temp, humidity, & light level should a At what temp, humidity, & light level should a

product be stored? In what form?: liquid, product be stored? In what form?: liquid, powder, ect.powder, ect.

6.1 Review Questions6.1 Review Questions

1.1. Why are antibiotics important Why are antibiotics important biotechnology products?biotechnology products?

Before the widespread use of Before the widespread use of antibiotics in the 1940’s many antibiotics in the 1940’s many people died from common diseases people died from common diseases that we now treat fairly easy such that we now treat fairly easy such as, strep throat, bronchitis, and as, strep throat, bronchitis, and pneumoniapneumonia

2.2. What is the function of the enzyme What is the function of the enzyme amylase?amylase?

Break down the large molecule starch Break down the large molecule starch into smaller molecules—glucose.into smaller molecules—glucose.

3. Why might a company be interested 3. Why might a company be interested in producing amylase as a product?in producing amylase as a product?

Since amylase may be used to produce Since amylase may be used to produce sugar, any industry needing large sugar, any industry needing large quantities of sugar would be quantities of sugar would be interested. It’s easy to produce and interested. It’s easy to produce and cheap!cheap!

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

See slide.See slide.

6.2 Review Questions6.2 Review Questions

1.1. What kind of assay would use Bradford What kind of assay would use Bradford reagent in the test?reagent in the test?

Protein concentration. It’s a nonspecific Protein concentration. It’s a nonspecific protein indictor and will show the protein indictor and will show the presence of any protein in solution. A presence of any protein in solution. A technician can determine the technician can determine the concentration of an unknown sample by concentration of an unknown sample by comparing the unknown to known comparing the unknown to known solutionssolutions

2. For what purpose would a 2. For what purpose would a technician use an ELISA?technician use an ELISA?

Determine the presence & Determine the presence & concentration of a specific protein concentration of a specific protein utilizing antibody-antigen utilizing antibody-antigen specificity.specificity.

3. What does a stability assay 3. What does a stability assay measure?measure?

Show the shelf life of a product. At Show the shelf life of a product. At what temperature, humidity, and what temperature, humidity, and light level should the product be light level should the product be stored. In what form should it be stored. In what form should it be stored: liquid, powder, freeze-dried, stored: liquid, powder, freeze-dried, capsules, etc.capsules, etc.

4. In a large company, which 4. In a large company, which department would have several department would have several employees developing and employees developing and conducting assays?conducting assays?

An entire department. Some An entire department. Some companies have Assay Services and companies have Assay Services and Quality Control Departments that Quality Control Departments that specialize in testing company specialize in testing company products.products.