sepharose and dansyl-chloride

In this report, we describe a strategy for enhancing the immunogenicity of a wide variety of Ags by linking them to IL-2 via an IgG3-IL-2 fusion protein with high affinity for a convenient hapten Ag, dansyl (DNS; N,N-dimethyl-1-aminonaphthalene-5-sulfonyl chloride). This fusion protein, anti-DNS-IgG3-IL-2, combines the functional characteristics of its constituents and has pharmacokinetic properties that are greatly improved over those of IL-2 and a previously described IgG1-IL-2 fusion. The molecule is intact and recoverable from the blood of mice hours after i.p. injection and reaches distant organs throughout the animal. The 7-h in vivo half-life of this molecule is much longer than that of IL-2, addressing a major obstacle in the application of IL-2 to human diseases, including cancer and AIDS. Additionally, the Ab's specificity for the hapten dansyl and the convenient chemistry of dansyl provide a means to link IL-2 to virtually any molecule of interest without the complexities and uncertainties of making IL-2 fusions with each molecule individually. Using hapten-conjugated-BSA (DNS-BSA) as a model Ag we show that the Ab response elicited by anti-DNS-IgG3-IL-2-bound DNS-BSA-Sepharose injected into mice is increased over that of DNS-BSA-Sepharose or anti-DNS-IgG3-bound DNS-BSA-Sepharose. Anti-DNS-IgG3-IL-2 also increased the Ab response to soluble DNS-BSA after a booster injection. This system should be useful in testing the ability of IL-2 to potentiate the immune response to Ag and in screening a large number of potential Ags for use in vaccines. The dramatically improved pharmacokinetics should also overcome one of the major difficulties in applying IL-2 to the treatment of human disease, its short half-life.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Dansyl Compounds; Female; Half-Life; Haptens; Immunization; Immunoglobulin G; Interleukin-2; Mice; Mice, Inbred BALB C; Recombinant Fusion Proteins; Sepharose; Serum Albumin, Bovine; Tissue Distribution

Topics: Dansyl Compounds; Enzymes, Immobilized; Indicators and Reagents; Methods; Microspheres; Sepharose; Sulfinic Acids

Topics: Chromatography, Affinity; Dansyl Compounds; Enzymes, Immobilized; Indicators and Reagents; p-Dimethylaminoazobenzene; Sepharose; Sulfinic Acids; Urease