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

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Articles

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