Department of Biological Engineering, Sichuan University of Science & Engineering, Zigong, China.
Department of Gastroenterology, FuShun People's Hospital, Zigong, China.
Int Dent J. 2024 Dec;74(6):1298-1305. doi: 10.1016/j.identj.2024.05.006. Epub 2024 May 25.
The purpose of this work was to develop an anti-CAT-SYI immunoglobulin Y (IgY) antibody that targeted both GtfB (glucosyltransferase B) and GbpB (glucan-binding protein B) and test its anticaries properties in rats.
A new CAT-SYI fusion gene was created utilising functional DNA fragments from the GtfB and GbpB genes. The recombinant antigens, comprising the fused CAT-SYI antigen, GtfB, and GbpB, were expressed and purified using a prokaryotic expression and purification system. The purified recombinant antigens were utilised to immunise laying hens against particular IgY production. The biological activities of these particular IgY antibodies were then assessed both in vitro and in vivo, including their capacity to suppress biofilm formation and tooth caries.
Results indicated that these produced IgY antibodies demonstrated a high antibody titer (>0.1 μg/mL) and could precisely recognise and bind to their respective antigens. Furthermore, it was discovered that these specific IgY antibodies successfully bind to Streptococcus mutans and significantly reduce biofilm development. After 8 weeks of ingesting antigen-specific IgY meals, comprising anti-GtfB IgY and anti-GbpB IgY, the severity of dental caries was dramatically reduced in S mutans-infected Sprague-Dawley rats (P < .01). Anti-CAT-SYI IgY therapy significantly reduced tooth cavities by 89.0% in vivo (P < .05) compared to other treatment groups.
The anti-CAT-SYI IgY, a multitarget antibody that targets both GtfB and GbpB, displayed excellent inhibitory effects against S mutans, making it a promising targeted method with improved anticaries efficacy and significant application opportunities.
本研究旨在制备靶向 GtfB(葡萄糖基转移酶 B)和 GbpB(葡聚糖结合蛋白 B)的 CAT-SYI 免疫球蛋白 Y(IgY)抗体,并在大鼠体内检测其抗龋性能。
利用 GtfB 和 GbpB 基因的功能 DNA 片段,构建了新的 CAT-SYI 融合基因。采用原核表达和纯化系统表达和纯化融合抗原,包括融合 CAT-SYI 抗原、GtfB 和 GbpB。用纯化的重组抗原免疫产蛋母鸡,以产生特异性 IgY。评估这些特异性 IgY 抗体的体外和体内生物活性,包括抑制生物膜形成和龋齿的能力。
结果表明,这些产生的 IgY 抗体表现出高抗体滴度(>0.1 μg/mL),能够精确识别和结合相应的抗原。此外,发现这些特异性 IgY 抗体能与变形链球菌结合,并显著减少生物膜的形成。在摄入含抗 GtfB IgY 和抗 GbpB IgY 的抗原特异性 IgY 饮食 8 周后,变形链球菌感染的 Sprague-Dawley 大鼠的龋齿严重程度显著降低(P <.01)。与其他治疗组相比,抗 CAT-SYI IgY 治疗组体内龋齿减少 89.0%(P <.05)。
靶向 GtfB 和 GbpB 的多靶点抗体抗 CAT-SYI IgY 对变形链球菌具有良好的抑制作用,是一种具有改善抗龋效果和广阔应用前景的有前途的靶向方法。