An exhaustive body of data has been generated that indicates the importance of efforts to further develop cyanovirin-N (CV-N) as an anti-HIV-1 microbicide. It is a highly potent inhibitor of cell-free and cell-to-cell transmission of HIV-1 infection, with no observable toxic effect on host cells. The primary obstacle to advancing CV-N as a microbicide is the fact that it is currently difficult to produce in quantities required for proper preclinical study and clinical evaluation. As a recombinant protein, CV-N must be produced by means of an in vivo recombinant expression system. Our central hypothesis in this Project is that being of prokaryotic origin, CV-N should be highly compatible with bacteria-based expression systems, and its unique biochemical properties make it feasible to apply an array of purification technologies that typically would not be appropriate in efforts to obtain a bioactive recombinant protein. Therefore, we have defined a set of Specific Aims that are designed to: (1) comprehensively study the regulation of CV-N expression in bacterial systems; (2) exploit the unique biochemical properties of CV-N in the development of a high yield purification scheme; (3) determine the relevance of specific growth parameters to the expression of CV-N in larger scale fermentation; and (4) test if alternative forms of CV-N can be engineered that either enhance production or HIV-1 inhibitory activity. These efforts will be achieved through extensive interaction between the research team of this project, and the other Research Projects and Cores of the Program.
|Su, H Irene; Schreiber, Courtney A; Fay, Courtney et al. (2011) Mucosal integrity and inflammatory markers in the female lower genital tract as potential screening tools for vaginal microbicides. Contraception 84:525-32|
|Barnhart, Kurt; Kulp, Jennifer L; Rosen, Mark et al. (2009) A randomized trial to determine the distribution of four topical gel formulations in the human vagina. Contraception 79:297-303|
|Owen, Derek H; Peters, Jennifer J; Kieweg, Sarah L et al. (2007) Biophysical analysis of prototype microbicidal gels. J Pharm Sci 96:661-9|
|McFadden, Karyn; Cocklin, Simon; Gopi, Hosahudya et al. (2007) A recombinant allosteric lectin antagonist of HIV-1 envelope gp120 interactions. Proteins 67:617-29|
|Tien, Deborah; Schnaare, Roger L; Kang, Feirong et al. (2005) In vitro and in vivo characterization of a potential universal placebo designed for use in vaginal microbicide clinical trials. AIDS Res Hum Retroviruses 21:845-53|
|Colleluori, Diana M; Tien, Deborah; Kang, Feirong et al. (2005) Expression, purification, and characterization of recombinant cyanovirin-N for vaginal anti-HIV microbicide development. Protein Expr Purif 39:229-36|