Production of a single-chain variable fraction capable of inhibiting the Streptococcus mutans glucosyltransferase in Bacillus brevis:: construction of a chimeric shuttle plasmid secreting its gene product
Production of a single-chain variable fraction capable of inhibiting the Streptococcus mutans glucosyltransferase in Bacillus brevis:: construction of a chimeric shuttle plasmid secreting its gene product
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DOI：
10.1016/s0167-4781(03)00038-1
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发表时间：
2003-04-15
期刊：
影响因子：
--
通讯作者：
Abiko, Y
中科院分区：
文献类型：
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作者：
Shiroza, T;Shinozaki-Kuwahara, N;Abiko, Y
牙周炎和龋齿是当今常见的口腔疾病，被动免疫是最有效的预防方法之一。为此，我们构建了小鼠和人单克隆抗体来抑制牙龈卟啉单胞菌相关的血凝和共聚集。此外，还构建了也抑制血凝的人工抗体、单链可变部分或scFv。特别针对龋齿，还构建了抑制负责生物膜形成的葡萄糖基转移酶（GTF）活性的小鼠和人类单克隆抗体。 scFv 相对于天然抗体的优势在于，前者分子不会诱导 Fc 可能产生的副作用，例如自身免疫性疾病，因为它仅由源自重链和轻链的可变区组成。为了提高抗体制剂的能力，我们尝试使用短芽孢杆菌（一种分泌能力强的革兰氏阳性细菌）作为宿主细胞构建额外的 scFv。通过采用新的异二聚体系统构建嵌合穿梭质粒，成功地从短双歧杆菌转化体中分泌出与亲本抗体具有相同生物活性的scFv蛋白。 (C) 2003 Elsevier Science B.V 保留所有权利。
Periodontitis and dental caries are common oral diseases, in these days, and the passive immunization is one of the most effective approaches for prevention. For this purpose, we have constructed mouse and human monoclonal antibodies to inhibit the Porphyromonas gingivalis-associated hemagglutination and coaggregation. In addition, an artificial antibody, single-chain variable fraction, or scFv, which also inhibited the hemagglutination, was constructed. Specifically for dental caries, mouse and human monoclonal antibodies that inhibited the glucosyltransferase (GTF) activity, responsible for biofilm formation, were also constructed. The advantage of scFv over the native antibody is that the former molecule does not induce possible side-effects due to Fc, such as autoimmune disease, because it consists only of variable regions originating from both heavy and light chains. To increase the abilities of the antibody preparations, we attempted to construct an additional scFv using Bacillus brevis, a secretion-proficient gram-positive bacterium, as a host cell. An scFv protein possessing the same biological activity as that of the parental antibody was successfully secreted from a B. brevis transformant following the construction of a chimeric shuttle plasmid, which was accomplished by employing a new heterodimer system. (C) 2003 Elsevier Science B.V All rights reserved.