Immunoglobulin

Question 1

Albumin

Answer 1

Most abundant plasma protein

Maintains osmotic pressure

Transports FAs and metals (has hydrophobic pockets and will bind many hydrophobic drugs)

Malnourished children will have edema in stomach and face if not making enough albumin

Question 2

Fibrinogen

Answer 2

Blood coagulation

Question 3

Epitope

Answer 3

Small specific portion of that protein that the antibody recognizes

Question 4

IgG

Answer 4

Most common

Seen in many therapeutic drugs

Heavy chain = gamma

Question 5

IgA

Answer 5

Tears, saliva, protects external body surfaces

Dimer

Mucosal surfaces

Alpha

Question 6

IgM

Answer 6

Produced early in immune response, and exists as a pentamer

Alot of binding sites

Use IgMs to see how long an antigen has been in your system

Mu

Question 7

IgD

Answer 7

Exist on lymphocytes

Unknown function

Delta

Question 8

IgE

Answer 8

Persist at low levels in serum, functional for parasitic reactions

You can have problems with allergeric reactions if don’t have enough IgE

Epsilon

Question 9

Heavy chains

Answer 9

Twice as long as light chains

Each Ig has two heavy chains connected by disulfide bonds

Question 10

Light chains

Answer 10

Either kappa of lambda

Never a mixture

Question 11

Beta-pleated sheets in Igs

Answer 11

H-bonds between different sequences of AAs

Resulting in a pleated structure

Important because the terminal AA will end on different levels which will fold up…

Creates a hydrophobic face and hydrophilic face

Question 12

‘Beta-sandwich’

Answer 12

Immunoglobulin fold

Two sheets come together

Greasy parts come inside

This is how Igs come together = supersecondary structure

Question 13

Variable regions

Answer 13

N-terminal region

Supersecondary structures - where antigen binds

Also known as = ‘Fab region’

Question 14

CDR

Answer 14

Complementary determining region

Where sheets are connected by loops

Where antigen binds

Regions of HYPERVARIABILITY….explains epitope diversity

Question 15

‘Fc Region’

Answer 15

Heavy chains not in the variable region are known as this….

Property to crystallize and control the effector portion of the Ig

Question 16

ELISA

Answer 16

1. Coat surface with antigen
2. Fill in the rest of the space with unspecified proteins that will not interact with the experiment at all
3. Incubate a primary antibody that is specific to the antigen
4. Incubate a secondary antibody that is specific to the primary antibody (with an enzyme bound to it)
5. Incubate substrate that enzyme has a affinity for
6. Have enzymatic product be fluorescent

Darker = more concentration of antigen..

Therefore ELISA allows for quantification

Question 17

Western blot

Answer 17

Proteins are separated by size in a denaturing SDS gel

Total proteins can be visualized by staining the gel with a dye that binds proteins

Compare to where bands show up in sample gel

Able to determine size of antigen

Question 18

ChIP assay

Answer 18

Investigate the interaction between protein and DNA in the cell

It can tell whether specific proteins are associated with specific regions in the genome

Useful to determine regions of genome that epigentic modifications occur

—> cross-link proteins to DNA…shearing the DNA into short pieces and then using antibodies to immunoprecipitate proteins from the complex mixture

Question 19

Polyvlonal antibodies

Answer 19

Your body will make a bunch of antibodies for the foreign protein in your body by B-cells

Not all of them will bind to an epitome …

So may have 100s of antibodies binding to other pieces

—> mixture of antibodies that recognize different epitomes

Question 20

Monoclonal antibodies

Answer 20

More homogenous

All antibodies recognize the same epitome

There are naturally occuring monoclonals antibodies

Question 21

Multiple Myeloma

Answer 21

B-cells expand and get large concentration of the IgGs in your serum

Patients have high M protein concentration in serum,

They secrete light chains in urine because they cant reabsorb all that was produced = BENCE JONES protein

Symptoms = elevated Ca2+ levels, renal failure, bone lesions (CRAB), and anemia

Question 22

ScFv

Answer 22

Binding activity is in the end termianl (SCFV)

First variable region of the light and heavy chains are linked through a peptide bond

This is a molecule that has thrown away everything except the light and heavy chain with N-terminal and the CDR

It still retains the binding properties of the Ig