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


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Authors

  • Snatashree Mohanty Central Institute of Freshwater Aquaculture
  • M. Makesh
  • K. V. Rajendran
  • P. P. Suresh Babu
  • Deepika Anand
  • Saurav Kumar
  • Abhay Kumar
  • Raju Baitha
  • Kamal Sarma

https://doi.org/10.21077/ijf.2019.67.2.84594-08

Abstract

Serum immunoglobulins  (Ig) of mrigal Cirrhinus mrigala (Hamilton 1822) immunised with bovine serum albumin (BSA), were purified by affinity chromatography using BSA-CL agarose column. The purified mrigal Ig (m-Ig) was characterised under reducing condition by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) which revealed two bands of 85 and 26 kDa corresponding to heavy and light chain, respectively. Following fusion of splenocytes from Balb/c mice immunised with purified m-Ig with myeloma cells, three hybridomas showing reactivity with m-Ig were cloned by limiting dilution. The monoclonal antibodies (MAbs) generated by these clones were designated as 3B2-E12, 3B2-F9 and 4C3-B2 and characterised by western blotting and isotyping. Western blot analysis of the supernatant from the three clones with purified m-Ig indicated that, all the three MAbs were specific to heavy chain. Isotyping revealed that 3B2-E12 MAb was of IgG1 isotype whereas the other two MAbs were of IgG2a isotype. Cross reactivity of anti-mrigal Ig MAb (3B2-E12) was observed with serum Ig of Catla catla and Labeo rohita indicating semi-conserved nature of Ig in Indian major carps.

Keywords: Cirrhinus mrigala, ELISA, Immunoglobulin, Monoclonal Antibodies, Western blotting

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Submitted

2019-01-05

Published

2020-06-30

Issue

Vol. 67 No. 2 (2020)

Section

Articles

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The copyright of the articles published in Indian Journal of Fisheries vests with the Indian Council of Agricultural Research, who has the right to enter into any agreement with any organization in India or abroad engaged in reprography, photocopying, storage and dissemination of information contained in these journals. The Council has no objection in using the material, provided the information is being utilized for academic purpose but not for commercial use. Due credit line should be given to the ICAR where information will be utilized.

How to Cite

Mohanty, S., Makesh, M., Rajendran, K. V., Suresh Babu, P. P., Anand, D., Kumar, S., Kumar, A., Baitha, R., & Sarma, K. (2020). Production and characterisation of monoclonal antibodies against immunoglobulins of Cirrhinus mrigala (Hamilton 1822). Indian Journal of Fisheries, 67(2). https://doi.org/10.21077/ijf.2019.67.2.84594-08