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Biochemistry Division
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Chemiluminescence is the generation of electromagnetic radiation as light energy from a chemical reaction. Light
emitting reactions which take place by the use of electrical current are called Electro-chemi-luminescent reactions.

Electro-chemi-luminescent (ECL) processes are known to occur with numerous molecules including compounds of
ruthenuim, osmium or other elements. This is a process in which highly reactive species are generated from stable
precursors of an electrode. These highly reactive species react with one another producing light. The development
of ECL immuno assay is based on the use of ruthenium (11) tris (bipyridyl) Ru (bpy3)2+complex and tripropylamine
(TPA). The final chemi-luminescent product is formed during the detective step.

The chemical reactions that lead to the emission of light from the ruthenium complex are initiated electrically rather
than chemically. This is achieved by applying a voltage to the immunological complexes (including the Ruthenium
complex) that are attached to streptavidin coated micro particles .The advantage of eclectically initiating the
chemiluminescent reaction is that the entire reaction can be precisely controlled. Two electrically active
substances, the ruthenium complex and the tripropylamine (TPA) are involved in the reactions that lead to the
emission of light. Both substances remain stable. This reaction occurs at the surface of the Platinum electrode. The
applied voltage creates an electrical field which causes all the materials in the field to react. The ruthenium ground
state complex is continuously regenerated. This complex can perform many light generating cycles during the
measurement process, therefore showing an inherent amplification effect which contributes to the technologies
sensitivity. Many protons can be created from one antigen antibody complex.

ECL SIGNAL GENERATION

The graph displays a typical ECL signal generation when a voltage is applied to the detection cell electrode; a
peak of light emission occurs over a short time interval and can be detected as the resulting ECL signal. A defined
area under the curve is measured around the intensity maximum.

ADVANTAGES OF ECL TECHNOLOGY

Extremely stable non-isotopic label allows liquid reagent convenience.
Enhanced sensitivity in combination with short incubation times means high quality assays and fast results turn
around.
Large measuring range minimizes dilutions and repeat, including handling time and reagent loss.

TEST PRINCIPLE:

1. Competitive principle
2. Sandwich principle
3. Bridging principle