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An Envelope Domain III–based Chimeric Antigen Produced in Pichia pastoris Elicits Neutralizing Antibodies Against All Four Dengue Virus Serotypes

There is currently no vaccine to prevent dengue (DEN) virus infection, which is caused by any one of four closely related serotypes, DEN-1, DEN-2, DEN-3, or DEN-4. A DEN vaccine must be tetravalent, because immunity to a single serotype does not offer cross-protection against the other serotypes. We have developed a novel tetravalent chimeric protein by fusing the receptor-binding envelope domain III (EDIII) of the four DEN virus serotypes. This protein was expressed in the yeast, Pichia pastoris, and purified to near homogeneity in high yields. Antibodies induced in mice by the tetravalent protein, formulated in different adjuvants, neutralized the infectivity of all four serotypes. This, coupled with the high expression potential of the P. pastoris system and easy one-step purification, makes the EDIII-based recombinant protein a potentially promising candidate for the development of a safe, efficacious, and inexpensive, tetravalent DEN vaccine.

Received June 6, 2007. Accepted for publication March 20, 2008.

Acknowledgments: The authors thank Dr. Andrew Falconar for providing all four serotypes of DEN viruses.

Financial support: This work was supported by ICGEB core funds and a grant from the Department of Biotechnology, Government of India, to SS and NK. BE and SK were supported during the course of this work by an ICGEB predoctoral fellowship and a CSIR (Government of India) senior research fellowship, respectively.

^* Address correspondence to Navin Khanna, RGP Group, PO Box 10504, International Centre for Genetic Engineering and Biotechnol-ogy, Aruna Asaf Ali Marg, New Delhi 110067, India. E-mail: navin{at}icgeb.res.in

Authors’ addresses: Behzad Etemad, Gaurav Batra, Rajendra Raut, Satinder Dahiya, Saima Khanam, Sathyamangalam Swaminathan, and Navin Khanna, RGP Group, PO Box 10504, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India, Tel: 91-11-26742357, Fax: 91-11-26742316.

Reprint requests: Navin Khanna, RGP Group, PO Box 10504, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India, Tel: 91-11-26742357, ext. 272, Fax: 91-11-26742316, E-mail: navin{at}icgeb.res.in.