The Selective Isolation And Quantitative Study Of Three Species Rid Tide Algaes By Immunology Technology

Rapidly and accurately detect and enumerate the red tide algaes species was one of research direction in harmful algal blooms. However, the present detecting and monitoring techniques are not adequately prepared to predict harmful algal blooms, As a consequence, a great deal of effort has been made to increase the sensitivity of the diagnostic process and to reduce the time spent to obtain a result.Polyclonal antibodies against three red tide algae with different size, Heterosigma akashiwo, Gymnodinium sp and Alexandrium catenella were raised in this study. Block affinity treatment was done to enhance the specificity of the antibodies. The titer of three polyclonal antibodies was determined by indirect enzyme-linked immunosorbent assay, and the specificity against polyclonal antibody was determined by indirect competitive enzyme-linked immunosorbent assy(icELISA). The result showed that Heterosigma akashiwo and Gymnodinium sp antibodies without block treatment got titer higher than 1:41 000 and 1:18 000, respectively, an indicating of high affinity antibodies. The titer of Alexandrium catenella antibody reached 1:3 500, relatively lower than the previous two. The specificity of three polyclonal antibodies after affinity treatment was raised remarkably. However, the titer declined. All the polyclonal antibodies showed a high affinity with their own cells. Polyclonal antibodies were not remarkable cross reacted with other algae in the icELISA.Distributing of antigen on the outer algae cells surface and the specificity of three polyclonal antibodies was researched by indircct Fluoroimmunoassay, rapidly and accurately detect and enumerate the red tide algaes cells by flow cytometry method. The results suggest that the fluorescent signal of Heterosigma akashiwo and Gymnodinium sp antibodies was conferred to the outer cell surface, while those of Alexandrium catenella was patchy distributed mainly around the excavated cingulum. Polyclonal antibodies were only reacted with their own algae. No cross activity with other two algae was detected. The antibody of Alexandrium catenella reacted to its own cells, no cross activity was found with Alexandrium tamarense, which will be applicable in distinguishing species within the genus. The results of flow cytometry method suggest that the otherness between the cell counts by light microscopy and the cell counts by flow cytometry method. The otherness in Heterosigma akashiwo is + 5.16%, The otherness in Symbiodinium sp is + 4.65%, The otherness in Alexandrium catenella is +11.76%. this method was rapidsensitive,steady and selective isolation,quantitative study of rid tide algaes, even if no taxology knowledge.the selective isolation and quantitative of Heterosigma akashiwo by immuno-magnetic bead separation (IMBS) method, the result suggest that Heterosigma akashiwo was separated by immuno-magnetic bead, the hallmark is 85%--95%, the percent of were capture is 60%--90%, this method can be used to the selective isolation and quantitative of algae cells and selective isolation live cells, this method was convenient and alterable.Phage Display Peptide Library was used to selective and isolate specificity peptide. specificity peptide was researched to antigen on the outer algae cells surface and selective isolation of Heterosigma akashiwo by this technology. A usful technology was provided to detect red tide alga.