The Preparation Of Polyclonal Antibodies Against Some Red Tide Algae, Development Of An Enzyme-linked Immunosorbent Assay And Its Application For Quantifications Of The Algae

In this study, polyclonal antibodies (PAbs) were respectively raised by immunizing rabbits with whole cells against nineteen harmful algae of China sea. After the analysis of their titer and specificity, several were selected to develop the indirect competitive enzyme-linked immunosorbent assay (icELISA). A good result was achieved with the method in detection of some simulating natural samples.After immunized by Chaetoceros curvisetus, the mice, hens and rabbits all produced PAbs against the alga, and different immunological procedures had little influence on the antibodies' producing and specificity. Less time was used for the rabbits to produce the PAbs with a higher titer and production, so the rabbit was chosen to produce PAbs in the subsequent experiments. All the PAbs showed a high affinity, with a titer of 6000 to 69000, most had cross activities with other algae, and only a few showed a good species-specificity. But the cross activities may be reduced greatly after properly adsorbed with other algae.The antisera against Chaetoceros debilis, Heterosigma akashiwo, Prorocentrum micans and Gymnodinium sanguineum were used to establish the icELISA respectively. The C. debilis antisera can recognize C. curvisetus, C. gracilis, C. minutissimus as well as C. debilis with a detect limit of about 51 cells in the icELISA. The other three antisera had good species-specificity, with a detect limit of several tens to several hundreds. The least detecting number of H. akashiwo and P. micans was about 77 and 55 cells correspondingly. Quantifications of some simulating samples were analyzed by the icELISA and the results coincided with those of manual counts approximately. In addition, a discuss about the use of indirect ELISA(iELISA) in the phytoplankton research was also made in the paper.