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Journal of Virology, January 2009, p. 98-104, Vol. 83, No. 1
0022-538X/09/$08.00+0     doi:10.1128/JVI.01564-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Design and Expression of a Dimeric Form of Human Immunodeficiency Virus Type 1 Antibody 2G12 with Increased Neutralization Potency {triangledown}

Anthony P. West Jr.,1 Rachel P. Galimidi,1 Christopher P. Foglesong,1 Priyanthi N. P. Gnanapragasam,1 Kathryn E. Huey-Tubman,1,2 Joshua S. Klein,1 Maria D. Suzuki,1 Noreen E. Tiangco,1 Jost Vielmetter,1,3 and Pamela J. Bjorkman1,2*

Division of Biology,1 Howard Hughes Medical Institute,2 Caltech Protein Expression Center, California Institute of Technology, 1200 E. California Blvd., Pasadena, California 911253

Received 24 July 2008/ Accepted 10 October 2008

The antigen-binding fragment of the broadly neutralizing human immunodeficiency virus type 1 (HIV-1) antibody 2G12 has an unusual three-dimensional (3D) domain-swapped structure with two aligned combining sites that facilitates recognition of its carbohydrate epitope on gp120. When expressed as an intact immunoglobulin G (IgG), 2G12 formed typical IgG monomers containing two combining sites and a small fraction of a higher-molecular-weight species, which showed a significant increase in neutralization potency (50- to 80-fold compared to 2G12 monomer) across a range of clade A and B strains of HIV-1. Here we show that the higher-molecular-weight species corresponds to a 2G12 dimer containing four combining sites and present a model for how intermolecular 3D domain swapping could create a 2G12 dimer. Based on the structural model for a 3D domain-swapped 2G12 dimer, we designed and tested a series of 2G12 mutants predicted to increase the ratio of 2G12 dimer to monomer. We report a mutation that effectively increases the 2G12 dimer/monomer ratio without decreasing the expression yield. Increasing the proportion of 2G12 dimer compared to monomer could lead to a more potent reagent for gene therapy or passive immunization.

* Corresponding author. Mailing address: Division of Biology, California Institute of Technology, 1200 E. California Blvd., Pasadena, CA 91125. Phone: (626) 395-8350. Fax: (626) 395-5939. E-mail: bjorkman{at}caltech.edu

{triangledown} Published ahead of print on 22 October 2008.

Journal of Virology, January 2009, p. 98-104, Vol. 83, No. 1
0022-538X/09/$08.00+0     doi:10.1128/JVI.01564-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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