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Biotechnology and Applied Biochemistry (2001) 34, (63–69) (Printed in Great Britain)

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Recombinant Opc protein from Neisseria meningitidis reconstituted into liposomes elicits opsonic antibodies following immunization

Tania Carmenate, Circe Mesa, Tamara Menéndez, Viviana Falcón and Alexis Musacchio¹

Division of Vaccines, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba

Key words: immune response, phospholipid vesicles, protein refolding, vaccines.

Abbreviations used: AEC, 3-amine-9-ethylcarbazole; BHI, Brain Heart Infusion; CTB, cholera toxin-B subunit; DRV, dried–reconstituted vesicle; HRP, horseradish peroxidase; LPS, lipopolysaccharide; mAbs, monoclonal antibodies; MLVs, multilamellar vesicles; OMPs, outer membrane proteins; PBST, PBS containing 0.05% Tween 20; rOpc, recombinant Opc.

¹To whom correspondence should be addressed (e-mail alexis.musacchio@cigb.edu.cu).

The reconstitution of recombinant bacterial outer membrane proteins (OMPs) into their native conformations after purification has been the major problem in their use as effective vaccines. Liposomes have been shown to be an attractive approach, providing a native-like environment for these antigens. The meningococcal recombinant Opc (rOpc) protein, produced as inclusion bodies in Escherichia coli, was incorporated into phospholipid vesicles consisting of dipalmitoyl phosphatidylcholine and cholesterol. The incorporation of rOpc into the lipid bilayer was demonstrated, and the reconstitution of some native epitopes was tested using a set of monoclonal antibodies. Subcutaneous immunization of Balb/c mice with rOpc-containing vesicles resulted in the generation of a high level of specific antibodies. The elicited antibodies reacted with the native meningococcal protein and showed opsonic activity.

Received 26 January 2001/26 April 2001; accepted 9 May 2001

Portland Press Ltd © 2001