Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

Jang, Han Na; Woo, Jong Kyu; Cho, Young Hye; Kyong, Seo Bong; Choi, Suna

doi:10.5483/bmbrep.2004.37.3.314

Cited by 17 publications

(14 citation statements)

References 17 publications

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“…Additional proteins in the vicinity of the 75 kDa major band were also observed in some aliquots eluted from the BSA-affinity column, or detected by western blot in the serum of several fish species used in this study. Evidence for the existence of distinct forms of H (Palenzuela et al 1996, Watts et al 2001, Jang et al 2004, Suresh Babu et al 2008) and L chain (Espelid & Grøntvedt 2003, Ishikawa et al 2004, Feng et al 2009) within teleost fish has already been reported and our data indicates that this could also be true for silver catfish and the other fish evaluated here. However, slight differences in the molecular weight of a protein might be attributed to different pattern of glycosylation.…”

Section: Discussionsupporting

confidence: 65%

Characterization of an IgM-like immunoglobulin from silver catfish (Rhamdia quelen) serum and its use for the production of polyclonal antibodies and development of immunoassays

Kreutz

Canova²,

Nied³

et al. 2016

Pesq. Vet. Bras.

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Knowledge on fish immunoglobulin (Ig) characteristics and the availability of monoclonal or polyclonal antibodies to fish Igs are essential to evaluate the humoral immune response and the Ig distribution on leukocyte cells. We demonstrated that silver catfish serum Ig is composed of one immunodominant H chain with approximately 75k Da and one L chain with approximately 28 kDa, similar to human IgM. Rabbit polyclonal antibodies to the catfish IgM-like Ig recognized both the H and L chain and were useful in developing an indirect ELISA to measure the production of antibodies in fish immunized with bovine serum albumin. Dot blot and western blot cross-reactivity studies indicated a wide degree of epitope sharing amongst Ig from several Siluriformes and Characiformes fish indigenous to Brazilian rivers. In these fish species, polyclonal antibodies reacted mostly with the H chain. The results presented here are central to the development of tools and strategies to investigate the antibody production to inoculated antigens and tissue distribution of Ig molecules in native fish species. Furthermore, because of the wide range of cross-reactivity, polyclonal antibodies to silver catfish IgM-like Ig might be used to develop immunoassays to measure the humoral immune response in other fish species.

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Section: Discussionsupporting

confidence: 65%

Characterization of an IgM-like immunoglobulin from silver catfish (Rhamdia quelen) serum and its use for the production of polyclonal antibodies and development of immunoassays

Kreutz

Canova²,

Nied³

et al. 2016

Pesq. Vet. Bras.

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“…In addition, B-cell heterogeneity has also been observed using monoclonal antibody produced against serum Ig in carp [15] and sea bass [12]. Compared to the reports of MAbs produced against Japanese flounder Ig previously [2][3][4], several differences and similarities were observed in the results reported here. In the present study, JFW20 and JFW21 reacted with 25.2-26.1 and 5.2-8.3% of the PBL, respectively.…”

Section: Discussioncontrasting

confidence: 49%

“…Western blotting showed that JFW20 recognized 70 and 74 kDa antigens, which corresponded to the heavy chains of Ig [3]. JFW21 did not react with reduced flounder Ig, but it did react with an 880 kDa (data not shown) nonreduced serum protein.…”

Section: Discussionmentioning

confidence: 70%

“…[4]. Jang et al found that 49% of the HKL was positive for anti-Japanese flounder Ig MAb [3]. While the underlying reasons for these differences may correspond to Ig heterogeneity in Japanese flounder, the influence of physiological or genetic differences should also be considered.…”

Section: Discussionmentioning

confidence: 95%

“…While Japanese flounder is frequently used for immunological studies, only monoclonal antibody (MAb) against immunoglobulin (Ig) has been reported for use as a cell surface marker for Ig ? lymphocytes [2][3][4]. Production of additional monoclonal antibodies for use as markers of leukocyte subpopulations is central to the further analysis of flounder immunology.…”

Section: Introductionmentioning

confidence: 99%

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Production and characterization of monoclonal antibodies to leukocytes and serum immunoglobulin in Japanese flounder Paralichthys olivaceus

et al. 2009

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Two monoclonal antibodies (MAbs: JFW1 and JFW10) were produced against peripheral blood leukocytes (PBL) in Japanese flounder. Additionally, MAbs against flounder immunoglobulin (Ig; JFW20 and JFW21) were generated for the surface marker of Ig ? leukocytes using purified serum Ig as an antigen. MAb JFW1 recognized the surface marker of granulocytes and monocytes and MAb JFW10 specifically bound to the surface antigen of thrombocytes. Flow cytometric analysis of PBL incubated with JFW1, JFW10, JFW20 and JFW21 revealed that 2.5-7.4, 23.7-50.1, 25.2-26.1 and 5.2-8.3% of all leukocytes were positive for these markers. Analysis of head kidney leukocytes (HKL) showed that JFW1, JFW10, JFW20 and JFW21 bound to 30.5-36.3, 1.9-2.8, 6.4-8.3 and 1.9-3.0% of all leukocytes, respectively. Western blot analysis after SDS-PAGE showed that JFW10 recognizes a protein of 115 kDa from lysed PBL. JFW20 recognized the 70 and 74 kDa proteins of the heavy chain of Ig from serum. No band was observed for either JFW1 or JFW21. These antibodies will be useful for the identification and isolation of Japanese flounder leukocyte subpopulations and will facilitate immunological studies of flounder.

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Monoclonal antibody-based immunodot for evaluating serum immunoglobulin levels in rohu (Labeo rohita)

Babu

Shankar²,

Honnananda

et al. 2014

Aquacult Int

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Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

Cited by 17 publications

References 17 publications

Characterization of an IgM-like immunoglobulin from silver catfish (Rhamdia quelen) serum and its use for the production of polyclonal antibodies and development of immunoassays

Characterization of an IgM-like immunoglobulin from silver catfish (Rhamdia quelen) serum and its use for the production of polyclonal antibodies and development of immunoassays

Production and characterization of monoclonal antibodies to leukocytes and serum immunoglobulin in Japanese flounder Paralichthys olivaceus

Monoclonal antibody-based immunodot for evaluating serum immunoglobulin levels in rohu (Labeo rohita)

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