short communication | DOI: https://doi.org/10.31579/2693-7247/157
Immunology of Invertebrates, Orléans University (France).
*Corresponding Author: Michel Leclerc, Immunology of Invertebrates, Orléans University (France).
Citation: Michel Leclerc, (2024), The Sea Star Igkappa Cloning by the use of e. Coli and Cho Protocol. J. Pharmaceutics and Pharmacology Research, 7(2); DOI:10.31579/2693-7247/157
Copyright: © 2024, Michel Leclerc. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received: 13 November 2023 | Accepted: 01 December 2023 | Published: 29 January 2024
Keywords: protocol; protein; production
This previous work has performed with UMR-INRA ISP1282 (Nicolas Aubrey). It allows, by the use of CHO (Chinese Hamster Ovarian) protocol cloning to produce in a relative great quantity the « Young Protein » or anti-HRP (Horse-Radish Peroxydase) from the sea star IGKappa gene
10 years ago, we tried to clone, for the first time, the sea star IG Kappa gene by the use and the help of E. coli as amplificator [1]. It allowed, in à second time, to verify that the Young Protein, or anti-HRP Protein recognizes the HRP antigen [2].
But, this verification of the affinity between the IPA (Invertebrate Primitive Antibody) and the antigen, seemed unclear at that time, for many of us. Thus, we decided to operate new cloning [3] with new parameters and new affinity tests.
The last of them, used CHO protocol, as described in various experiments [4].
First, the percentage of Young Protein production, was greatly ameliorated with the CHO protocol. A best rate occurs when compared to the E. coli one sensu stricto. A western-blot determined exactly the M.W of the Young Protein, as shown in Figure 1: It is 12,49271 Kda.
Figure 1 : Purification of Young Protein (Production en ExpiCHO)
We think now to perform an Elisa test to verify the affinity between this young protein and the HRP antigen.
We envisage also (My colleagues and me) in a next future, to immunize other sea stars with anti-tumoral antigens to product specific nanobodies (Ref.4) from sea stars, against cancer activity (in a general way): sequencing and cloning, after’’ll be applicated to obtain a specific recombinant specific protein.
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