ELISA

Título: Técnica de ELISA

Miembros: Sabrina Santos, Miranda Guerra
Obje1vos:
- Conocer más a fondo acerca de los an>cuerpos y las dis>ntas técnicas moleculares
u>lizadas para iden>ficarlos.
- Llevar a cabo de manera eficaz un ELISA
Anotaciones:
- ELISA
o Detec>ng or quan>fying
o
o
Immunology
- An>gen: any substance that induces immune
response (usually proteins)
- An>body: immune system proteins that recognize
an>gens via affinity bindings
o Structure:

ANTIGEN DETECTION ELISA COMPONENTS

- ANTIGEN: pa>ent sample is tested for
presence of an>gens from viruses, bacteria,
etc.
- Primary an1body:
o Binds to an>gen.
o Can be produced in lab by injec>ng
target an>gen into animal and then
harves>ng the serum.
- Secondary an1body
o Binds to constant region of primary
an>body.
o Made by injec>ng primary an>bodies
from one animal to a different
animal.
- Substrate
o Changes color in presence of enzyme = indicates posi>ve
 ELISA
- Virus detec>on at protein level
- Based on immune system an>body molecules
Type Descrip>on Advantages Disadvantages Figure
1. Add an>gen Simplest and Least sensi>ve
2. Primary AB fastest
Direct

3. Add chromogenic
substrate

1. Add an>gen Cheaper and Longer protocol

2. 1ry AB more sensi>ve
a. Binds to
Indirect

an>gen
3. 2ry AB
a. Binds to
1ry AB
4. Add substrate
1. Coat microplate Highly sensi>ve Complex to
with capture AB op>mize AB cross-
2. Add an>gens that reac>vity between
affinity capture and
a. bind to detec>on AB
capture AB
3. Add detec>ng AB
a. Bind to
an>gen
4. Add Enzyme-linked
Sandwich

2ry AB
a. Binds to
detec>ng
AB
5. Add substrate
a. Reacts with
enzyme-
linked
secondary
AB for
an>gen
detec>on
Compe>t
ive
 Immune System

Immune System Descrip1on An1body Produc1on

Innate Immunity Acts instantly upon pathogen
detec>on.
Most common an>body: IgM
General response

Adap>ve immunity Takes hours to days to develop.

Most common an>body: IgG
Highly specific response
Creates immune memory.
B-cell and T-cell dependent

Infec1on 1meline

ELISA + COVID-19
- Spike protein targe>ng by Elisa.
 o ACE2 receptor recogni>on and host
invasion
§ ACE2 is found in lung, heart,
intes>ne, and liver >ssues.
o TMPRSS2 recognizes S protein,
facilita>ng viral invasion.
- Spike protein
o S1 subunits- ACE 2
§ RBD
• Recognizes ACE2
receptor
§ NTD
o S2 subunits- membrane fusion
§ 6 helical bundle via two-heptad repeat domain (HR1, HR2)

- COVID-19 medica1on
o Inhibi>ng ACE2 or TMPRSS22 recogni>on
§ Treats COVID-19 infec>ons
- COVID ELISA detec1on
o Nucleocapsid
§ (N, binds the RNA genome)
o Membrane (M)
o Envelope (E)
o Spike (S)