Learn: Capstone Lab Quiz 1

SOP means

Standard Operating Procedures (SOPs)

What does an SOP tell you?

- Procedures for dealing with spills, emergencies, and exposure incidents

- Methods for limiting employee exposure (PPE, etc.)

Is risk assessment qualitative or quantitative?

Qualitative

What are some factors we use for Risk Assessment?

- Pathogenicity

- Route of Transmission

- Agent Stability

- Infectious Dose

- Concentration

Pathogenicity

ability of a microorganism to cause disease

Route of Transmission

the anatomical and physiological path of transmission from an infected source to a human

Infectious Dose

the number of microorganisms or viruses needed to establish an infection

What do we do with risk assessment information?

Agents are split into four risk groups based upon their relative pathogenicity to healthy adult humans

Risk Group 1

Agents are not associated with disease in healthy adult humans

Risk Group 2

Agents are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available.

Risk Group 3

Agents are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available.

Risk Group 4

Agents are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available.

Primary Containment

the protection of personnel and the immediate laboratory environment from exposure

Who does primary containment protect?

YOU (the researcher)

What are some ways we practice primary containment?

- Good microbiological technique

- Use of appropriate safety equipment

- PPE

Secondary Containment

the protection of the environment external to the laboratory from exposure

What are some ways we practice secondary containment?

- Facility design

- Operational practices

BSL-1 Labs are usually used for

- basic teaching

- research

BSL-2 Labs are usually used for

- primary health services

- diagnostic services

- research

BSL-3 Labs usually used for

- special diagnostic services

- research

GMT

good microbiological techniques

BSL-? --> GMT only

BSL-1

BSL-? --> GMT, PPE, biohazard sign

BSL-2

BSL-? --> Special clothing, controlled access, directional air flow

BSL-3

BSL-? --> Air lock entry, shower exit, special waste disposal

BSL-4

What BSL level is our capstone lab?

BSL-2

Wear PPE, Lab Coat, Gloves

You may not need protective eyewear

BSL?

BSL-1

- Eye protection is required, it's no longer optional

- Face/eye protection

- PPE is mandatory

- Has additional safety equipment (hoods, biosafety cabinets)

- Will include biosafety bins and infectious hazard bins

BSL?

BSL-2

- Has an ante-room (room you step into before entering the lab)

- Negative air flow

- Air's being pulled into the room, so if the air is contaminated, it can't slip out the door into the hallways

- Autoclave within the lab

BSL?

BSL-3

- Few select labs in the state/country

- Completely isolated environment

- Everything's completely sterile and separate

- Requires a UV room to sterilize

BSL?

BSL-4

Orange Bag

Biohazard Bag

Red Bag

Infectious Waste Bag

Can sharps go in the orange bags?

NO

Once an orange bag is treated and labeled, where can it go?

Into normal trash

What do infectious waste bags look like?

- Red color

- Tear/leak resistant

- Have universal biohazard symbol

What must be kept for all chemicals?

MSDS

MSDS =

Material Safety Data Sheet

Where are our MSDS?

In binders at the front of the room, which includes MSDS, SOP (Material Safety Data Sheet)

How can a biohazard bag go into the normal trash?

- Sterilize them in the autoclave for at least 90 minutes at 250 degrees F

- Once it comes out of the autoclave and it's sterilized, it can go into the regular trash can

By whom are professional standards of conduct/ethics created by?

They are developed by professional communities themselves for themselves --> doctors, lawyers, engineers, etc.

In order to do funded scientific research in the United States, is ethics training required?

Yes, they are required courses that one must take

What are some examples of violations of scientific integrity?

- Copying

- Sharing

- Stealing

- Lying

- Plagarism

What is plagirism?

The act of presenting another person's work or ideas as your own

What are some ways that scientists can go wrong when conducting experiements?

- Honest error in measuring, recording, and interpreting

- Lack of review

- Overinterpretation

- Lack of reproducibility

How can scientists keep themselves on the right track?

- Double and triple checking data

- Mentorship

- Having their work reviewed by senior authors

What are some consequences of when scientists go wrong?

- Loss of credibility

- Retraction of published work

- Loss of degree/job

Where do antibodies come from?

The immune systems of living systems

Immunology

The study of the structures of the immune systems

What is the adaptive immune response?

- Reacts to antigens

- Remembers an invader

- Highly specific to antigen

- Takes time to acquire

What do B cells do?

Produce antibodies in the bone marrow

What is the innate immune response?

- Requires no prior contact (unlike adaptive)

- Involves inflammatory process

What do antibodies do?

- Bind to antigens on pathogens

- Seek to disable the pathogen's activity

What is an antigen?

- Not part of the host

- Large polysaccharide or protein that lies on the surface of viruses and foreign cells

What 2 things trigger active immunity

Infection or vaccination

B cells are involved in what type of immune response?

Humoral immune response

T cells do what

Attack cells infected w/bacteria or viruses

T cells are involved in what type of immune response?

Cell-mediated immune response

What is an epitope?

The specific region on antigen to which the antibody binds

Epitope

What shape does an antibody take on?

Y-shape

How many antigens can bind to a single epitope?

1 epitope is capable of binding only 1 antigen --> each antigen has a specific location

Antibody

Where is the constant region of an antibody?

E

Where is the constant region of an antibody?

At the bottom of the Y-shape of the antibody

Why are antibodies big business?

Because they are used in so many ways, which include --> purification, protein visualization, and SDS-page

What are the big 3 applications of antibodies which make them "big business"

- Purification

- Protein visualization

- SDS-page

How do we make antibodies?

- In the lab

- By injecting an animal with an antigen

- This leads to the formation of antibodies

What are polyclonal antibodies?

A mixture of antibodies produced that recognize several different epitopes on an antigen

How are polyclonal antibodies made?

--> An antigen (e.g. protein) is injected into an animal

--> A mixture of antibodies is produced and isolated

--> Each antibody in the mixture recognizes a different, specific epitope within the protein.

What's cheaper/easier to produce, polyclonal or monoclonal antibodies?

Polyclonal antibodies

What are monoclonal antibodies?

Identical antibodies that are specific to the specific epitope of a protein

How are monoclonal antibodies made?

- Start with B cells

- Put through cell culture

- Decide which cell makes 1 kind of antibody

- Multiply that cell

- Create a cell culture that produces this singular antibody

Which provides stronger data, monoclonal or polyclonal antibodies?

Monoclonal

Which provides more specific data, monoclonal or polyclonal antibodies?

Monoclonal

What is the most promising class of drugs?

Monoclonal antibodies

If a drug ends in "-mab" what does that mean?

- It's expensive

- Highly efficacious

ELISA stands for

Enzyme Linked Immunosorbent Assay

What 2 types of antibodies do we use for ELISA testing?

- Primary antibodies

- Secondary antibodies

What is a primary antibody in ELISA testing?

They bind to the antigen in our sample, but we can't see it b/c it's so small

What is a secondary antibody in ELISA testing?

It binds to the primary antibody to see if the primary antibody is found on the constant region

Constant Region

Bottom portion of the Y

In ELISA testing, if the antigen is in our sample what happens?

- The primary and secondary antibodies bind

- A color change occurs

What causes/catalyzes a color change in ELISA testing?

- Enzyme

- Found in secondary antibody

What has an enzyme (used for possible color change catalysis)--the primary or secondary antibody?

Secondary

What is the drawback of ELISA testing?

- Not super sensitive

- Doesn't work instantly (can change over time)

Are antibodies sticky? Why is this important?

- Yes they're very sticky

- This enables them to stick to test tubes, blood, etc.

Why are blocking reagents used in ELISA testing?

- To fill in sticky sites

- To prevent non-specific binding

--> We want the antibody to stick to COVID and not the test tube

What does a positive control do?

It makes sure the test is working, should turn the positive result of the test

What can cause a false positive

- Pipetting error

- Contamination

- Recent vaccination

What is a false positive

Occurs when an assay determines a "positive" result for a target substance/disease that is not actually present in the sample

--> can be caused by cross-reactivity, sample handling, or technical errors

What is a false negative?

It occurs when a test determines that a substance/condition is not present when it actually is

What can cause a false negative

--> Testing the patient too early, they haven't been sick long enough to make antibodies

--> Switching patient samples

What type of assay does a pregnancy test use?

ELISA