Biomedical Engineering Reference
In-Depth Information
Target protein
Antibody
coated well
Target protein
binding to
the antibody
ds DNA-linked
target specific
antibody binding
to target
PCR amplification of the
label DNA and
detection of fluorescence signal
4.6
Quantitative immuno PCR (qIPCR) mechanism.
4.5.1 Immuno-PCR assay
Quantitative immuno-PCR (qIPCR) is a highly sensitive immunoassay
that combines the advantage of protein quantifi cation by ELISA and the
power of signal amplifi cation by PCR (Fig. 4.6). This novel assay is capa-
ble of robust detection of very low concentration of proteins with specifi c
antibodies conjugated with double-stranded DNA as a signal molecule that
can be amplifi ed by PCR (Saiki
et al.
, 1988). Signals amplifi ed with increased
DNA products can be detected by real-time PCR. Quantitative IPCR has
been shown to be 100-10 000 times more sensitive than a conventional
ELISA, and can be used to detect trace proteins in a sample that may not
be detectable by other conventional immunoassays (Niemeyer
et al.
, 2005;
Sano
et al.
, 1992). Recently, a few commercial qIPCR kits have been devel-
oped for resolving the challenges and it is possible that this assay will be
widely used for protein analysis in the future.
In summary, many techniques have been presented in this chapter as a
guide for cell differentiation analysis. The strategies adopted for character-
ization will depend on the quality of information that is most relevant to
the needs of the researcher. Many of these techniques are well established
and will require the researcher to have intimate understanding of specifi c
biological pathways and systems. This information is readily available in the
research literature and the references provided in this chapter. The impact
of the technologies in molecular biology and imaging analysis have been
adapted to characterize biomaterials and monitor the success of basic tis-
sue engineering research. In the future, adaptation and development of new
methods will lead to increased throughput of biomaterials and characteriza-
tion of their three-dimensional structures.
