E3 Journal of Biotechnology and Pharmaceutical Research
E3 Journal of Biotechnology and Pharmaceutical Research Vol. 2 (1) pp. 001-008 , October 2011; © E3 Journals; ISSN 2141-7474
Isolation and binding affinity of C-phycocyanin to blood cells and genomic DNA as well as its diagnostic applicationsPoonam Singh , Mohammed Kuddus * , George Thomas
Jacob School of Biotechnology and Bioengineering, SHIATS, Allahabad, India.
*Corresponding Author E-mail: email@example.com
Accepted 10 September 2011
In the analytical field, many of the chemicals used as fluorescent probes are dangerous and expensive. The use of natural flourochromes like C-phycocyanin (C-PC) in diagnostics is being looked upon as a replacement for organic flourochromes. In this study, C-PC was extracted from algal biomass by six different methods, and analyzed for its binding affinity towards red blood cells of human and fowl blood, platelets and lymphocytes of humans, and genomic DNA of human and plant. The comparative staining ability of C-PC and the commonly used ethidium bromide stain towards the genomic DNA was finally analyzed. The freezing and thawing method resulted in a maximum C-PC yield and the algal biomass dried under shadow by air circulation gave the higher purity. The maximum binding affinity of C-PC was observed with human lymphocytes at 25Ã‚Â°C and a C-PC dilution ratio of 1:104. In case of fowl red blood cells, the maximum affinity was observed at 25Ã‚Â°C with dilution ratio of 1:105. The comparative study on the staining ability of ethidium bromide and C-PC towards the genomic DNA revealed a more or less similar strength of reaction. Thus, it is proposed that the partially purified natural dye, C-PC, could be used as a substitute of ethidium bromide. It may be applied as a fluorescent reagent for immunological analysis.
Keywords: C-phycocyanin, blood cells, DNA staining, fluorescent microscopy, Spirulina.[Download Article - PDF]