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Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii)

Fish and Shellfish Immunology

By:
, , , , , , and
DOI: 10.1016/j.fsi.2012.06.006

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Abstract

Pacific herring (Clupea pallasii) have a central role in the North Pacific ecosystem as a forage fish species and are natural reservoirs of several important finfish pathogens, including Viral hemorrhagic septicemia virus (VHSV). Here, we report the identification of the gene encoding the immunoglobulin mu (IgM) heavy chain, as well as the development and characterization of monoclonal antibodies (MAbs) that specifically react with Pacific herring IgM. Pacific herring immunoglobulin was purified and consisted of heavy and light chains of approximately 80 and 25 kDa. Three hybridoma clones were initially identified by ELISA as reactive with purified immunoglobulin but only one clone was able to detect an 80 kDa protein in Pacific and Atlantic herring (Clupea harengus) whole plasma by denaturing western blot. However, all three MAbs were able to precipitate an 80 kDa protein from Pacific herring and LCMS sequencing of peptide fragments derived from this protein matched the predicted amino acid sequence of the cloned, heavy chain gene. In addition, two of the MAbs stained cells within the putative lymphocyte gates for the spleen, anterior kidney and posterior kidney but were not reactive for myeloid/granulocyte gates, which is consistent with these MAbs reacting with surface IgM+ B-cells. To our knowledge, this is the first report of IgM-related gene sequences and anti-IgM monoclonal antibodies from any member of the family Clupeidae. The antibodies produced in this study are critical for achieving our long-term goal of conducting serological surveillance to assess pathogen exposure in natural populations of Pacific herring.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
Production and characterization of monoclonal antibodies to IgM of Pacific herring (Clupea pallasii)
Series title:
Fish and Shellfish Immunology
DOI:
10.1016/j.fsi.2012.06.006
Volume
33
Issue:
3
Year Published:
2012
Language:
English
Publisher:
Elsevier
Publisher location:
Amsterdam, Netherlands
Contributing office(s):
Western Fisheries Research Center
Description:
7 p.
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
Larger Work Title:
Fish and Shellfish Immunology
First page:
552
Last page:
558
Number of Pages:
8