Chemiluminescence and Bioluminescence, Analysis by
527
Fig. 3
Electrochemiluminescent
ruthenium(II)
tris(bipyridyl)–labeled DNA.
N
N
N
N
N
N
Ru
+ +
(CH
2
)
3
CO-DNA
Light
Tripropylamine
Electrode surface
Tab. 2
Illustrative examples of applications of chemiluminescent and
bioluminescent reactions.
Type of application or test
Examples
Antibodies
IgA, IgG, IgE, IgM
Antioxidants
Vitamin E
Biomass
Bacteriuria testing, enumeration of bacteria in milk,
soil biomass, shelf-life dating, sperm viability
Chemosensitivity
Biocide assessment, tumor chemosensitivity
Drugs
Digoxin, methotrexate
Environmental
Phenolic herbicides, hygiene monitoring
Enzymes
Adenylate kinase,
β
-1,4-galactosyltransferase,
creatine kinase, galactose 1-phosphate,
N
-acetyl-
β
-d-glucosaminidase, proteases
Gases
N
itr
icox
ide
,n
itrogend
iox
ide
,ozone
,su
lfurd
iox
ide
Hormones
Follitropin, lutropin, thyrotropin, thyroxine,
parathyrin
Lighting
Emergency and warning light, aerosols, and smoke
Metals
Ag, Bi, Ca, Ce, Co, Cr, Fe, Hg, K, Li, Mg, Mn, Ni,
Os,Pb
,Tb
,Th,T
l,V
,Zr
Oxidants
Hypobromite, hypochlorite, iodine, superoxide
Steroids
Estradiol, estriol, progesterone, testosterone
Substrates and cofactors
ADP, ATP, bile acids, ethanol, galactose, glucose,
glucose 6-phosphate, glycerol, lactate, NADH,
NADPH, oxaloacetate, phosphocreatine,
pyruvate, sorbitol
the reaction be initiated in front of the
photodetector using an injector. The wave-
length of light emission is a further
factor in selecting a detector. Photomul-
tiplier tubes have a wavelength-dependent
quantum efFciency for light detection
and usually are most sensitive in the
blue region (350–400 nm) and least sen-
sitive in the red region (600 nm) of the
spectrum.
4.2
Immunoassay, Blotting, and Nucleic Acid
Hybridization Assays
The
extreme
sensitivity
of
CL
and
BL
assays
(detection
limits
in
the
attomole–zeptomole range: 10
18
–10
21
mol)
has
made
them
ideal
for
immunoassay
and
other
applications
(Table 3). CL and BL substances are
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