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Control of antigen-binding to aluminum adjuvants and the immune response with a novel phosphonate linker

机译：用新型膦酸酯连接子控制抗原与铝佐剂的结合和免疫反应

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摘要

The strongest mechanism for adsorption of antigens to aluminum adjuvants is ligand exchange, which involves the replacement of a surface hydroxyl on the adjuvant by a terminal phosphate group of the antigen. A novel phosphonate linker was developed that allows the addition of phosphonate (C-PO3) groups to proteins under controlled and chemically mild conditions. Increasing the number of linkers per protein molecule progressively increased the adsorption strength to aluminum hydroxide adjuvant (AH) as measured by elution in serum. The effect of phosphonate conjugation on the antibody response was determined with hen egg lysozyme (HEL), a protein that has the same charge as AH at neutral pH and does not adsorb to AH. The phosphonylated form of HEL (HEL-P) adsorbed to AH, indicating that the ligand exchange interaction could overcome the electrostatic repulsion. Mice injected with HEL-P/AH had a higher antibody titer to HEL than mice injected with HEL/AH, especially at lower antigen doses, suggesting that adsorption of antigen has a dose-sparing effect. Conjugation of CRM197, an antigen that adsorbs electrostatically to AH, with phosphonate linkers did not enhance the antibody response, indicating that adsorption by either electrostatic or ligand exchange to AH is sufficient to enhance the antibody response. (C) 2013 Elsevier Ltd. All rights reserved.
机译：抗原吸附至铝佐剂的最强机制是配体交换，这涉及用抗原的末端磷酸基团取代佐剂上的表面羟基。开发了一种新颖的膦酸酯连接子，可在受控的化学温和条件下将膦酸酯（C-PO3）基团添加到蛋白质上。如通过在血清中洗脱所测量，增加每个蛋白质分子的接头数目逐渐增加了对氢氧化铝佐剂（AH）的吸附强度。用鸡蛋溶菌酶（HEL）测定了膦酸酯结合对抗体反应的影响，鸡蛋溶菌酶是一种在中性pH下与AH具有相同电荷且不吸附于AH的蛋白质。 HEL（HEL-P）的膦酰化形式吸附到AH上，表明配体交换相互作用可以克服静电排斥。注射HEL-P / AH的小鼠比注射HEL / AH的小鼠对HEL的抗体效价更高，尤其是在抗原剂量较低的情况下，这表明抗原的吸附具有一定的量效。 CRM197（一种静电吸附到AH的抗原）与膦酸酯连接物的结合不能增强抗体应答，表明通过静电或配体交换吸附到AH的吸附足以增强抗体应答。 （C）2013 Elsevier Ltd.保留所有权利。

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