Lecture 6 ELISA!. E L I S A nzyme- inked mmuno- orbent ssay Protein that facilitates a chemical...
-
Upload
darrell-bennett -
Category
Documents
-
view
213 -
download
0
Transcript of Lecture 6 ELISA!. E L I S A nzyme- inked mmuno- orbent ssay Protein that facilitates a chemical...
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