Production and Testing of Antibodies for Dreissena Mussels
Project ID: 9640
Principal Investigator: Kevin Kelly
Research Topic: Invasive Species
Priority Area Assignments: 2011 (Zebra and Quagga Mussels), 2012 (Zebra and Quagga Mussels), 2013 (Zebra and Quagga Mussels), 2014 (Zebra and Quagga Mussels)
Funded Fiscal Years: 2011, 2012, 2013 and 2014
Can we produce and isolate primary antibodies that can be used for the control, capture, and monitoring of Dreissena mussels?
Need and Benefit
During a mussel research meeting held on February 24, 2010, with invited scientists from the Metropolitan Water District of Southern California (MWD), Water Research Foundation (WaterRF), and the Bureau of Reclamation (Reclamation), one of the needs recognized during the meeting was to develop the use of antibodies specific for quagga and zebra mussels. As a result, one of the action items identified was to make a proposal for Dreissena antibodies to the Science and Technology (S&T) Program in collaboration with MWD. In April, a full proposal was submitted to the S&T Program with more details than what can be submitted online here.
In summary, there are multiple applications involving host-specific Dreissena antibodies that would benefit Reclamation. This would be the focus of the second year of this project, after candidate primary antibodies are screened and isolated during the first year.
1. Immunomagnetic Capture. Secondary antibodies used as part of a capture system are commercially available. For example, magnetic beads coated with antimouse antibodies can be used to capture Dreissena veligers that have been coated with mouse anti-Dreissena monoclonal antibodies. Similar capture systems such as this have been developed and are quite common. Such a capture system may be used either in the field or in the laboratory to selectively capture veligers from a background of other plankton species.
2. Immunofluorescent Detection. Secondary antibodies labeled with fluorescent probes are very useful for the selective detection of Dreissena veligers under the microscope. The immunodetection method is easier and faster than traditional microscopy methods. In contrast to the experience required for a microscopist to identify larvae on the basis of morphological traits (which are greatly lacking in bivalve larvae), immunodetection does not require any previous experience. An inexperienced person can easily identify green and non-green larvae after one teaching session. Identification of Dreissena veligers becomes much simpler, even with samples containing a large amount of plankton material. During a recent robin blind sample study, all laboratory methods, including microscopy, had associated errors that were greatest at the lowest larvae concentration despite the fact that these blind samples were prepared in very clean water samples at near zero plankton background levels.
3. Cell Counting. The need to analyze individual veligers under a microscope makes it difficult to study a large number of Dreissena larvae and many samples at the same time. Immunolabeling of veligers in the same way as for immunofluorescent detection (#2 above) makes it possible to automate veliger counting efficiently and accurately via conventional flow cytometry.
4. Conjugated Antibodies. Conjugated antibodies may be used as toxin delivery vehicles similar to human monoclonal antibody therapy for serious diseases such as multiple sclerosis and different types of cancers. The toxin can be attached to the antibodies that target specific cell types, such as cells found only within quagga and zebra mussels. These conjugates may be used as homing devices to deliver the toxin directly to the target species or even more narrowly to the target tissue cells where quagga and zebra mussels are most susceptible. Release of these conjugates at low levels in the water would have no effect, and only when these conjugates are concentrated enough on the target cells do they reach the minimum dosage level required to cause mortality.
Bureau of Reclamation Review
The following documents were reviewed by experts in fields relating to this project's study and findings. The results were determined to be achieved using valid means.
Monoclonal Antibodies for Improved Detection of Quagga Mussel Larvae (interim, PDF,
By Kevin Kelly
Research Product completed on May 18, 2015
Monoclonal Antibodies for Improved Detection of Quagga Mussel Larvae (final, PDF,
By Kevin Kelly
Research Product completed on May 19, 2015