Lecture 6

ELISA!

ELISA

nzyme-

inked

mmuno-

orbent

ssay

Protein that facilitates a chemical reaction

Immune system - antibodies

Survey technique:• High throughput• Overview

Antibodies?

VirusNH3

+

OH

O-

O

O

S

O

-O

O

+H3N

H-bonding signature

Antibodies?

N

N

N

Cl

NH

NH

Atrazine

How can you use antibodies to detect it?

Antibodies?

N

N

N

S

NH

NH

ProteinExpose

Collect sera

Harvest atrazine specific

antibodies

How ELISA works

Competition for the antibody active sites between:

• Analyte of interest - Antigen (ie. Atrazine)• Analyte mimic or antigen mimic

The Major Players

N

N

N

Cl

NH

NH

Compound of interest

Atrazine

=

The Major PlayersCompetitor

N

N

N

Cl

NH

NH

Enzyme= E

ELISA

The Major PlayersThe Antibody

ELISA

Fe

Detection

H2N NH2 NHHNE

H2O2

Compound that is

detected

Blue Yellow

via enzyme catalyzed reaction

Cross-reactivity

N

N

N

Cl

NH

NH

N

N

N

Cl

NH

NH

N

N

N

Cl

H2N NH

N

N

N

S

NH

NH

Atrazine Desethyl Atrazine

Simazine Ametryn