The importance of surface charge in the optimization of antigen-adjuvant interactions.
The adsorptive behavior of the recombinant malarial antigens R32tet32, R32NS181 and NS181V20 to aluminum hydroxide and aluminum phosphate gels was studied as a function of pH and buffer ions. The Plasmodium falciparum antigen, R32NS181, and the P. vivax antigen, NS181V20, with isoelectric points (pI) of 5.9 and 5.5, respectively, adsorbed readily to the positively charged boehmite form of aluminum hydroxide gel. These two antigens displayed reversible, linear adsorption behavior in the pH range 5-9, with maximal adsorption observed at the lowest pH studied. The addition of acetate buffer ions had little effect on adsorption, while the presence of phosphate decreased adsorption for R32NS181 and NS181V20 by 25 and 40% respectively. The adsorptive behavior of these two antigens with the negatively charged adjuvant, aluminum phosphate, was markedly decreased. The converse situation was observed with the R32tet32 antigen, whose pI is estimated to be 12.8. There was minimal interaction of this antigen with aluminum hydroxide gel except in the presence of phosphate counter ions and significant, nonreversible adsorption with aluminum phosphate gel. Enhanced adsorption of R32tet32 to aluminum hydroxide gel in the presence of phosphate is suggested to be the result of a covalent bond between a surface aluminum and a phosphate anion that modifies the surface charge of the aluminum hydroxide gel. These results indicate that the role of complementary surface charges, both for the ionization state of the protein and for the aluminum adjuvants, is the key in optimizing conditions for significant antigen-adjuvant interactions.
Department of Pharmaceutical Research, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406-0939.
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D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000327:Adsorption; D000535:Aluminum; D017607:Aluminum Compounds; D000536:Aluminum Hydroxide / Q000737:chemistry; D000941:Antigens / Q000737:chemistry*; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007525:Isoelectric Focusing; D008024:Ligands; D010710:Phosphates; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D013499:Surface Properties; D014961:X-Ray Diffraction
D000276:Adjuvants, Immunologic; D017607:Aluminum Compounds; D000941:Antigens; D000953:Antigens, Protozoan; D007477:Ions; D008024:Ligands; D010710:Phosphates; D011994:Recombinant Proteins; D000536:Aluminum Hydroxide; D000535:Aluminum; C012714:aluminum phosphate