24 Diagnostic Applications of Phycobiliproteins
601
RT-PCR labeled with both digoxigenin and biotin during amplification was done.
Subsequently, amplicons were simultaneously bound to anti-digoxigenin microparticles and fluorescently labeled with streptavidin-R-phycoerythrin (Wedemeyer et al.
2000).
Aberrant expression of various antigens by leukemic blasts or lymphoma cells
was assessed performing fluorescent assay using monoclonal antibodies conjugated
with phycoerythrin (Gratama et al. 1998). Detection of apoptosis in cultured cells
was performed by four-color flow cytometric assay. In which, the nucleated cells
were selected with CD45-phycoerythrin conjugates. Further, detection of lymphocytes, apoptosis and distinguishing early and late apoptosis were performed using
other dyes (Hasper et al. 2000). Quantifying melanosome transfer from melanocytes
to keratinocytes and surface of skin was investigated using phycoerythrin as a tool. It
employed double label with FITC-conjugated antibody against the melanosomespecific protein gp100, and with phycoerythrin-conjugated antibody against the
keratinocyte-specific marker cytokeratin (Ma et al. 2010). Detection of intracellular signaling of cytokines by dimerization of interleukin, IL-7 receptor could be
detected using specific antibodies conjugated with phycoerythrin (Andersen et al.
2013). Measurement of intracellular signaling proteins using calibrated standardized
phycoerythrin-conjugated beads was performed which had potential applications in
monitoring efficacy of targeted therapy (Chang et al. 2010).
It is interesting to note that applicability of phycobiliproteins is expanded to the
environmental issues too. Detection of diarrhetic shellfish poisoning toxin, Okadaic
acid (OA) produced by toxigenic dinoflagellates was performed using magnetic beads
coated with streptavidin. It prevents hazard to public health (Pan et al. 2018). Using
R-phycoerythrin-AgNPs (silver nanoparticle) construct, detection of Cu
2+ ions based
on the quenching effect between R-PE-AgNPs and Cu
2+ ions, in combination with
the fluorimetric method was made (Xu et al. 2019).
In the case of patents, the diagnostic potentials of PBPs were exploited in the
investigation of proteins and proteomics, genomics, transgenomics, microbial detection, food safety, and environmental aspects (Table 24.9). For example, studies pertinent to the interaction of proteins, detection of enzymes and receptors, nucleic acid
sequences, genetic polymorphism, analysis of transgenes in plants, authentication
of microbial detection, and analysis of food contaminants were patented wherein
phycobiliprotein fluorescence detection systems were coupled.
24.2.10 Development of Methods of Using PBPs
as a Detection Component
of Diagnostic/Prognostic/Theranostic Products
Though the aforesaid research literature and patents also include the methods of
using phycobiliproteins in each one of the applications, there are exclusive work
and patents on the development of methods in terms of usage of phycobiliproteins,
601
RT-PCR labeled with both digoxigenin and biotin during amplification was done.
Subsequently, amplicons were simultaneously bound to anti-digoxigenin microparticles and fluorescently labeled with streptavidin-R-phycoerythrin (Wedemeyer et al.
2000).
Aberrant expression of various antigens by leukemic blasts or lymphoma cells
was assessed performing fluorescent assay using monoclonal antibodies conjugated
with phycoerythrin (Gratama et al. 1998). Detection of apoptosis in cultured cells
was performed by four-color flow cytometric assay. In which, the nucleated cells
were selected with CD45-phycoerythrin conjugates. Further, detection of lymphocytes, apoptosis and distinguishing early and late apoptosis were performed using
other dyes (Hasper et al. 2000). Quantifying melanosome transfer from melanocytes
to keratinocytes and surface of skin was investigated using phycoerythrin as a tool. It
employed double label with FITC-conjugated antibody against the melanosomespecific protein gp100, and with phycoerythrin-conjugated antibody against the
keratinocyte-specific marker cytokeratin (Ma et al. 2010). Detection of intracellular signaling of cytokines by dimerization of interleukin, IL-7 receptor could be
detected using specific antibodies conjugated with phycoerythrin (Andersen et al.
2013). Measurement of intracellular signaling proteins using calibrated standardized
phycoerythrin-conjugated beads was performed which had potential applications in
monitoring efficacy of targeted therapy (Chang et al. 2010).
It is interesting to note that applicability of phycobiliproteins is expanded to the
environmental issues too. Detection of diarrhetic shellfish poisoning toxin, Okadaic
acid (OA) produced by toxigenic dinoflagellates was performed using magnetic beads
coated with streptavidin. It prevents hazard to public health (Pan et al. 2018). Using
R-phycoerythrin-AgNPs (silver nanoparticle) construct, detection of Cu
2+ ions based
on the quenching effect between R-PE-AgNPs and Cu
2+ ions, in combination with
the fluorimetric method was made (Xu et al. 2019).
In the case of patents, the diagnostic potentials of PBPs were exploited in the
investigation of proteins and proteomics, genomics, transgenomics, microbial detection, food safety, and environmental aspects (Table 24.9). For example, studies pertinent to the interaction of proteins, detection of enzymes and receptors, nucleic acid
sequences, genetic polymorphism, analysis of transgenes in plants, authentication
of microbial detection, and analysis of food contaminants were patented wherein
phycobiliprotein fluorescence detection systems were coupled.
24.2.10 Development of Methods of Using PBPs
as a Detection Component
of Diagnostic/Prognostic/Theranostic Products
Though the aforesaid research literature and patents also include the methods of
using phycobiliproteins in each one of the applications, there are exclusive work
and patents on the development of methods in terms of usage of phycobiliproteins,