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| Preparation and Characterization of Polyclonal Antibody Against CD133 Derived from Camel |
| ZHAI Tian-tian, MA Xiao-ling, Muyassar, M LI Ling-xia, LI Jiang-wei |
| Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China |
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Abstract Objective: The purpose is confirming and acquiring of high titer anti-CD133 antibodies from immunized camel, and providing the basis for preparing anti-CD133 nanobody. Methods: The gene segment coding of CD133 extracellular region were PCR amplified and ligated into pET28a plasmid and construction of a prokaryotic expression vector. The recombinant CD133 protein was expressed by IPTG induction and purified with Ni-affinity chromatography. A male Bactrian camel and two New Zealand rabbits were immunized with purified rCD133 antigen respectively. The enzyme-linked immunosorbent (ELISA) and Western blotting were applied to assay the titer of polyclonal antibody and the specific binding to CD133 protein. Results: the ELISA results revealed that the titer of anti-CD133 antibody raised from camel reached about 1:106 after the 5th immunization, and the titer of anti-CD133 antibody raised from rabbit reached about 1:5×105 after the 4th immunization.The anti-CD133 antisera could bind rCD133 specifically in Western blotting. Conclusion: The high titer anti-CD133 polyclonal antibodies were raised in camel compared with rabbit and it lay a good foundation for the future experiment.
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Received: 18 June 2014
Published: 25 August 2014
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