Time Resolved Amplified Cryptate

Emission (TRACE) Technology
Principle
Non-radiative energy transfer from the donor to the acceptor, which is part of a chemically
modified, light-collecting algal protein, is the basis of the TRACE technology. This transfer occurs
between 2 fluorescent tracers: the donor (europium cryptate) and the acceptor (XL665).  The
result will show in a ratio of the energy emitted at 665 nm to that emitted at 620 nm (internal
reference) is calculated for each sample. In this method, signal intensity is proportional to the
number of antigen-antibody complexes formed, and therefore to antigen concentration. The
process will be further discuss next slide

ProCT becomes detectable within 2 to 4 hours after a triggering event and peaks by 12 to 24
hours.

On the one hand, the spectral overlap between donor emission and acceptor absorption spectra,
intensifies the cryptate's fluorescent signal and, on the other hand, increases the life cycle of the
acceptor signal, allowing the temporally delayed fluorescence to be measured. TRACE guarantees
that by removing unbound and undefined signals, only signs of interest are measured.

PCT and TRACE

At 337 nm, a nitrogen laser is aimed at a sample containing PCT and 2 fluorescently labeled
antibodies that identify various PCT peptide epitopes. The resulting signal is amplified at 665 nm
when both molecules are brought into close proximity by binding to PCT, and extended to last for a
few micro-seconds. This extension ensures that the signal can be measured after the fluorescence of
the background (common in biological samples) has decayed.

The donor molecule is a polyclonal sheep antibody labeled Europium Cryptate that recognizes
epitopes in the immature CT region, while the acceptor molecule is a monoclonal antibody labeled
XL665 that is raised against the PCT CCP-1 region.

Benefits

 Precise results
 Less waste
 Faster results
 No wash and separation step – because some test includes washing for the other molecules
won’t interfere to the results. This makes the complex between PCT and the donor-acceptor
complex more precise and fast results
 Homogenous immunoassay - Homogenous immunoassay it is when an PCT in the complex
of donor-acceptor is then measured . It is easier and faster to perform. Because it does not
require separation of molecules.