Inhibition of biofilm formation by monoclonal antibodies against Staphylococcus epidermidis RP62A accumulation-associated protein
Inhibition of biofilm formation by monoclonal antibodies against Staphylococcus epidermidis RP62A accumulation-associated protein
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DOI：
10.1128/cdli.12.1.93-100.2005
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发表时间：
2005-01-01
期刊：
影响因子：
--
通讯作者：
Bryers, JD
中科院分区：
文献类型：
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作者：
Sun, DQ;Accavitti, MA;Bryers, JD
表皮葡萄球菌表达一种140 kDa的细胞壁结合蛋白积累相关蛋白(AAP)，以生物膜的形式附着和积累在表面。用单抗(MAb)潜在地阻断AAP可以减少或消除生物医学设备中表皮葡萄球菌的细菌定植。在这里，我们报告了我们的努力，(I)分离AAP，(Ii)产生抗AAP的单抗，以及(Iii)确定单抗抑制表皮葡萄球菌生物被膜形成的有效性。一个M7表皮葡萄球菌突变体，据报道缺乏AAP表达，作为阴性对照。免疫后，含抗AAP多克隆抗体的小鼠血清可使表皮葡萄球菌生物被膜减少54%。精选的抗AAP单抗对表皮葡萄球菌的减少率不超过66%。与单抗相比，12C6/12A1和3C1/12A1两种单抗对表皮葡萄球菌的抑制率分别高达79%和87%。与以前的报道相反，生物膜缺陷的表皮葡萄球菌突变体M7在其细胞壁上表达了一种200 kDa的蛋白，该蛋白特异性地结合了AAP单抗。用微毛细管反相高压液相色谱-纳米电喷雾串联质谱仪对该M7蛋白进行多肽分析，结果表明该蛋白与AAP的同源性为53%。正在进行的研究将阐明AAP和M7200-kDa蛋白的动态表达，以确定它们在生物膜形成中的作用。
Staphylococcus epidermidis expresses a 140-kDa cell wall-bound protein accumulation-associated protein (AAP) to adhere to and accumulate as a biofilm on a surface. Potentially blocking AAP with a monoclonal antibody (MAb) could reduce or eliminate S. epidermidis bacterial colonization of biomedical devices. Here, we report on our efforts to (i) isolate AAP, (ii) generate MAbs against AAP, and (iii) determine the efficacy of MAbs to inhibit S. epidermidis biofilm formation. An M7 S. epidermidis mutant, reportedly deficient in AAP expression, was used as a negative control. Postinoculation murine sera, containing polyclonal antibodies against AAP, were able to reduce S. epidermidis biofilm formation by 54%. Select MAbs against AAP were able to reduce S. epidermidis by no more than 66%. Two MAb mixtures, 12C6/12A1 and 3C1/12A1, reduced S. epidermidis accumulation up to 79 and 87%, respectively, significantly more than individual MAbs. Contrary to a previous report, biofilm-deficient S. epidermidis mutant M7 expressed a 200-kDa protein on its cell wall that specifically bound AAP MAbs. Peptide characterization of this M7 protein by microcapillary reversed-phase high-pressure liquid chromatography-nanoelectrospray tandem mass spectrometry resulted in 53% homology with AAP. Ongoing studies will elucidate the dynamic expression of AAP and the M7 200-kDa protein in order to define their roles in biofilm formation.