Engineering lysine‐rich caleosins as carrier proteins to render biotin as a hapten on artificial oil bodies for antibody production |
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| Authors: | Ting‐Hang Liu Chia‐Lin Chyan Feng‐Yin Li Ying‐Jie Chen Jason T C Tzen |
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| Affiliation: | 1. Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan;2. Dept. of Chemistry, National Dong Hwa University, Hualien, Taiwan;3. Dept. of Chemistry, National Chung Hsing University, Taichung, Taiwan;4. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan;5. School of Chinese Medicine, China Medical University, Taichung, Taiwan |
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| Abstract: | It has been demonstrated that caleosin alone is sufficient to stabilize artificial oil bodies. A series of recombinant caleosins, mutated with 3, 5, 8, 11, 13, 15, and 17 extra Lys residues and over‐expressed in Escherichia coli, were used as carrier proteins to render biotin as a hapten on the surface of artificial oil bodies for antibody production. Biotinylation levels of the recombinant caleosins were step‐wisely elevated as the number of extra Lys residues increased, and the biotinylated Lys residues were identified by mass spectrometric analysis. Polyclonal antibodies against biotin were successfully generated in rats injected with artificial oil bodies constituted with each of the biotinylated caleosins. Moreover, those generated via the biotinylated caleosins with eight or more extra Lys residues no longer recognized caleosin. It appears that engineered Lys‐rich caleosins are suitable carrier proteins for the production of antibodies against small molecules. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011 |
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| Keywords: | antibody production artificial oil bodies biotin caleosin hapten |
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