Lunin V G, Sharapova N E, Tikhonova T V, Poletaeva N N, Galushkina Z M, Aksenova E I, Grabko V I, Velikodvorskaia G A, Lavrova N V, Anan'ina Iu V
Mol Gen Mikrobiol Virusol. 2009(1):21-7.
Development of new technology allows different antigens of a necessary degree of cleanliness to be obtained. This development is a major problem of modern medical biotechnology. A promising approach to this problem includes use of the affinity domains (tags) incorporated in structure of a recombinant antigen and capable to bind to corresponding sorbents. The method of preparation of ready-for-use injections containing complexes formed by soluble antigens on insoluble cellulose immunosorbent (not chemical conjugates) in one stage is based on the fusion protein technology. This approach includes preparation of two-component recombinant proteins containing an antigen of interest and the cellulose-binding domain (CBD), which spontaneously binds to cellulose containing sorbents with high binding constant. Research into the immunogenic properties of the CBD in the complex with cellulose and in the preparation of recombinant CBD in a rat model was performed. The titers of specific antibodies in rat serum induced by recombinant CBD and CBD in the complex with cellulose was evaluated. The CBD in the complex with cellulose was more immunogenic in comparison with CBD alone. The spectrum and levels of cytokines in collected rat serum induced by developed preparations was also measured using the microsphere-based Luminex Flowmetrix system (BioPlex). It was found that the amorphous cellulose was not an immunotolerant sorbent, because it induced the expression of the proinfammatory cytokines in vivo.
新技术的发展使得能够获得具有必要清洁度的不同抗原。这一发展是现代医学生物技术的一个主要问题。解决这个问题的一个有前景的方法包括使用并入重组抗原结构中且能够与相应吸附剂结合的亲和结构域（标签）。基于融合蛋白技术的一种方法是制备即用型注射剂，该注射剂包含由可溶性抗原与不溶性纤维素免疫吸附剂（而非化学偶联物）在一个阶段形成的复合物。这种方法包括制备包含感兴趣抗原和纤维素结合结构域（CBD）的双组分重组蛋白，该结构域以高结合常数自发地与含纤维素的吸附剂结合。对大鼠模型中CBD与纤维素复合物的免疫原性特性以及重组CBD的制备进行了研究。评估了重组CBD和CBD与纤维素复合物诱导的大鼠血清中特异性抗体的滴度。与单独的CBD相比，CBD与纤维素的复合物具有更强的免疫原性。还使用基于微球的Luminex Flowmetrix系统（BioPlex）测量了所开发制剂诱导的收集大鼠血清中细胞因子的谱和水平。发现无定形纤维素不是一种免疫耐受吸附剂，因为它在体内诱导促炎细胞因子的表达。
