Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae |
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| Authors: | Jian Huang Ruobing Wen Zhenmin Bao Zhenghong Sui Ningbo Sun Kyoungho Kang |
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| Institution: | HUANG Jian 1, WEN Ruobing 2, BAO Zhenmin 1, SUI Zhenghong 1, **, SUN Ningbo 3, KANG Kyoungho 4 1 Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao 266003, China 2 North China Sea Environmental Monitoring Center, State Oceanic Administration, Qingdao 266033, China 3 The Fifth Middle School of Zibo, Zibo 255000, China 4 Department of Aquaculture, Chonnam National University, Yeosu Jeonnam 550-749, Korea |
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| Abstract: | Over the last several decades, harmful algal blooms (HABs) have become a serious environmental problem in many parts of the
world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that
can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella, Thalassiosira sp., Cryptomonas sp., Alexandrium tamarense and Symbiodinium sp.), were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against
H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and
applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%–91.6% of the cells were captured from unialgal culture by IMBs, and only
5.3%–12.5% of the four other HAB microalgae species were captured, indicating that the constructed IMBs combined specifically
with the H. akashiwo cells. At the same time, flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique, 91.7% of the targeted cells were
sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and
antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques, especially
immuno-magnetic separation, have low associated cost, and are fast and simple processes compared with other techniques currently
in use. |
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| Keywords: | Heterosigma akashiwo polyclonal antibody immuno-magnetic beads FITC flow cytometry |
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