Production and characterisation of monoclonal antibodies against immunoglobulins of Cirrhinus mrigala (Hamilton 1822)

Mohanty, Snatashree; Makesh, M.; Rajendran, K. V.; Babu, P P Suresh; Anand, Deepika; Kumar, Saurav; Kumar, Abhay; Baitha, Raju; Sarma, Kamal

doi:10.21077/ijf.2019.67.2.84594-08

Cited by 3 publications

(2 citation statements)

References 29 publications

(52 reference statements)

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“…Hybridoma clones secreting monoclonal antibodies (MAbs) against mrigal IgM, developed at Aquatic Animal Health laboratory of Central Institute of Fisheries Education, were used for secondary antibody production (Mohanty, ). A cryopreserved hybridoma clone (Cm4A11) was thawed quickly in a water bath (37°C) and washed once with Iscove's Modified Dulbecco's Medium (IMDM) by centrifuging at 500 g for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Specific immune response in mucosal and systemic compartments of Cirrhinus mrigala vaccinated against Edwardsiella tarda: In vivo kinetics using different antigen delivery routes

Qadiri

Makesh

Rajendran

et al. 2018

J World Aquaculture Soc

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Mucosal immune barriers confer protection against invading fish pathogens. Here, we conducted an experiment for 60 days to assess the mucosal and systemic immune response in Mrigal (Cirrhinus mrigala), an Indian major carp. Fish were immunized with inactivated Edwardsiella tarda by four different routes, namely, oral, immersion, injection, and anal intubation. An indirect enzyme‐linked immunosorbent assay (ELISA) was used to measure the specific immune response (antibody) in serum and mucus (collected from skin, gill, and gut) of the fish on 0, 15, 30, 45, and 60 days postimmunization. For specific immune response in the serum, significantly higher (p < 0.05) optical density (OD) values were obtained in the anal group (0.52 ± 0.03) and in the oral group (0.48 ± 0.03). In the skin mucus, significantly higher OD values were obtained in the oral group (0.48 ± 0.04) and immersion group (0.32 ± 0.03). In the gill mucus, significantly higher OD values were obtained in the oral group (0.82 ± 0.08) and the immersion group (0.73 ± 0.03). In the gut mucus, significantly higher OD values were obtained in the immersion group (0.080 ± 0.007) compared to the rest of the treatments. Fish from all the groups were challenged with LD50 dose of E. tarda at the end of the experiment. We conclude that oral and immersion immunization routes offer better protection of C. mrigala compared to other antigen delivery routes.

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Section: Methodsmentioning

confidence: 99%

Specific immune response in mucosal and systemic compartments of Cirrhinus mrigala vaccinated against Edwardsiella tarda: In vivo kinetics using different antigen delivery routes

Qadiri

Makesh

Rajendran

et al. 2018

J World Aquaculture Soc

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“…Those methods include enzyme linked immunosorbent assay (ELISA) and immunoblotting procedures, flow cytometry (FCM) analysis, stereotyping and analysis of antigens in microbial pathogens in both, medical and veterinary sciences, as well as to diagnostic and therapeutic options for disease monitoring and treatment [1] . MAbs specifically reacting with immunoglobulins (Igs) provide effective tools for assessing antibody production following infection or vaccination, and for immunoblotting analysis of antigens recognized by the host immune system [2] .…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy

Velázquez¹,

Cruz²,

Pérez-Bernal³

et al. 2023

Fish and Shellfish Immunology Reports

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Potential Applications of Fluorescence-Activated Cell Sorting (FACS) and Droplet-Based Microfluidics in Promoting the Discovery of Specific Antibodies for Characterizations of Fish Immune Cells

et al. 2021

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Akin to their mammalian counterparts, teleost fish possess a complex assortment of highly specialized immune cells that are capable of unleashing potent innate immune responses to eradicate or mitigate incoming pathogens, and also differentiate into memory lymphocytes to provide long-term protection. Investigations into specific roles and functions of fish immune cells depend on the precise separation of each cell type. Commonly used techniques, for example, density gradient centrifugation, rely on immune cells to have differing sizes or densities and thus fail to separate between similar cell types (e.g. T and B lymphocytes). Furthermore, a continuously growing database of teleost genomic information has revealed an inventory of cellular markers, indicating the possible presence of immune cell subsets in teleost fish. This further complicates the interpretation of results if subsets of immune cells are not properly separated. Consequently, monoclonal antibodies (mAbs) against specific cellular markers are required to precisely identify and separate novel subsets of immune cells in fish. In the field of fish immunology, mAbs are largely generated using the hybridoma technology, resulting in the development of mAbs against specific cellular markers in different fish species. Nevertheless, this technology suffers from being labour-intensive, time-consuming and most importantly, the inevitable loss of diversities of antibodies during the fusion of antibody-expressing B lymphocytes and myeloma cells. In light of this, the focus of this review is to discuss the potential applications of fluorescence-activated cell sorting and droplet-based microfluidics, two emerging technologies capable of screening and identifying antigen-specific B lymphocytes in a high-throughput manner, in promoting the development of valuable reagents for fish immunology studies. Our main goal is to encourage the incorporation of alternative technologies into the field of fish immunology to promote the production of specific antibodies in a high-throughput and cost-effective way, which could better allow for the precise separation of fish immune cells and also facilitate the identification of novel immune cell subsets in teleost fish.

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Production and characterisation of monoclonal antibodies against immunoglobulins of Cirrhinus mrigala (Hamilton 1822)

Cited by 3 publications

References 29 publications

Specific immune response in mucosal and systemic compartments of Cirrhinus mrigala vaccinated against Edwardsiella tarda: In vivo kinetics using different antigen delivery routes

Specific immune response in mucosal and systemic compartments of Cirrhinus mrigala vaccinated against Edwardsiella tarda: In vivo kinetics using different antigen delivery routes

Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy

Potential Applications of Fluorescence-Activated Cell Sorting (FACS) and Droplet-Based Microfluidics in Promoting the Discovery of Specific Antibodies for Characterizations of Fish Immune Cells

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