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聚电解质复合物中对17β-雌二醇的免疫反应:杂交瘤克隆的抗原特异性和亲和力

Immune response to 17 beta-estradiol in polyelectrolyte complex: antigen specificity and affinity of hybridoma clones.

作者信息

Başalp A, Bermek E, Cirakoğlu B, Coka V, Mustafaev M I, Saraç A S

机构信息

Institute for Genetic Engineering and Biotechnology, Tübitak Marmara Research Center, Kocaeli, Turkey.

出版信息

Hybridoma. 1996 Jun;15(3):233-8. doi: 10.1089/hyb.1996.15.233.

DOI:10.1089/hyb.1996.15.233
PMID:8823622
Abstract

Complex formation between synthetic polyelectrolytes (PE) [poly-4-vinyl-N-ethyl (cetyl), pyridine bromides-PVP(R2, R16)], bovine serum albumin (BSA), or 17 beta-estradiol-BSA conjugate (BSA.E) was studied in neutral water. Weakly water-soluble (colloidal) complex was formed upon addition of BSA.E to PVP (R2, R16) solution at pH 7. A nonrandom distribution of the protein molecules between the coils of polycations and self-assembly in the nonstochiometric polycomplex particles took place. The immunogenic properties of PVP (R2, R16)-BSA.E polycomplex were investigated and the specificities of produced antibodies analyzed. 17 beta-Estradiol introduced in polyelectrolyte complexes (PE-BSA) was found to invoke considerable increases in the steroid-specific immunoresponse. However, a comparative study of immunogenic activity of polycomplexes versus BSA.E+incomplete Freund's adjuvant (IFA) mixtures revealed some differences in regards to the specificity of antibody production. In contrast to IFA+BSA.E systems, polycomplexes were able to generate estradiol-as well as BSA-specific antibodies. Such a carrier-directed response may be determined by increase in immunogenicity of weak antigenic determinants and/or by the exposure of internally located determinants upon complex formation with polyelectrolytes. Fusions following the two different immunization procedures resulted in the growth of comparable numbers of estradiol-specific monoclonal antibodies with apparently similar antigen affinities. Thus, immunizations using antigens in PEC appear to provide an efficient alternative to IFA.

摘要

在中性水中研究了合成聚电解质(PE)[聚-4-乙烯基-N-乙基(十六烷基)吡啶溴化物-PVP(R2,R16)]、牛血清白蛋白(BSA)或17β-雌二醇-BSA缀合物(BSA.E)之间的复合物形成。在pH 7时,将BSA.E添加到PVP(R2,R16)溶液中会形成弱水溶性(胶体)复合物。蛋白质分子在聚阳离子的线圈之间呈非随机分布,并在非化学计量的多聚复合物颗粒中进行自组装。研究了PVP(R2,R16)-BSA.E多聚复合物的免疫原性,并分析了产生的抗体的特异性。发现引入聚电解质复合物(PE-BSA)中的17β-雌二醇会导致类固醇特异性免疫反应显著增加。然而,对多聚复合物与BSA.E+不完全弗氏佐剂(IFA)混合物的免疫原活性的比较研究揭示了在抗体产生特异性方面的一些差异。与IFA+BSA.E系统不同,多聚复合物能够产生雌二醇特异性抗体以及BSA特异性抗体。这种载体导向的反应可能是由弱抗原决定簇免疫原性的增加和/或与聚电解质形成复合物时内部定位决定簇的暴露所决定的。两种不同免疫程序后的融合产生了数量相当的具有明显相似抗原亲和力的雌二醇特异性单克隆抗体。因此,使用PEC中的抗原进行免疫似乎提供了一种有效的替代IFA的方法。

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