Green Brett J, Nayak Ajay P, Lemons Angela R, Rittenour William R, Hettick Justin M, Beezhold Donald H
Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health , Centers for Disease Control and Prevention, Morgantown, West Virginia.
Monoclon Antib Immunodiagn Immunother. 2014 Dec;33(6):428-37. doi: 10.1089/mab.2014.0042.
Chaetomium globosum is a hydrophilic fungal species and a contaminant of water-damaged building materials in North America. Methods to detect Chaetomium species include subjective identification of ascospores, viable culture, or molecular-based detection methods. In this study, we describe the production and initial characterization of a monoclonal antibody (MAb) for C. globosum enolase. MAb 1C7, a murine IgG1 isotype MAb, was produced and reacted with recombinant C. globosum enolase (rCgEno) in an enzyme-linked immunosorbent assay and with a putative C. globosum enolase in a Western blot. Epitope mapping showed MAb 1C7 specific reactivity to an enolase decapeptide, LTYEELANLY, that is highly conserved within the fungal class Sordariomycetes. Cross-reactivity studies showed MAb 1C7 reactivity to C. atrobrunneum but not C. indicum. MAb 1C7 did not react with enolase from Aspergillus fumigatus, which is divergent in only two amino acids within this epitope. The results of this study suggest potential utility of MAb 1C7 in Western blot applications for the detection of Chaetomium and other Sordariomycetes species.
球毛壳菌是一种亲水真菌,也是北美受水损害建筑材料的污染物。检测球毛壳菌属物种的方法包括子囊孢子的主观鉴定、活菌培养或基于分子的检测方法。在本研究中,我们描述了一种针对球毛壳菌烯醇酶的单克隆抗体(MAb)的制备及其初步特性。制备了鼠IgG1同种型单克隆抗体1C7,并在酶联免疫吸附测定中与重组球毛壳菌烯醇酶(rCgEno)反应,在蛋白质免疫印迹中与一种假定的球毛壳菌烯醇酶反应。表位作图显示单克隆抗体1C7对一种烯醇酶十肽LTYEELANLY具有特异性反应,该十肽在粪壳菌纲真菌中高度保守。交叉反应性研究表明单克隆抗体1C7与黑褐毛壳菌有反应,但与印度毛壳菌无反应。单克隆抗体1C7与烟曲霉的烯醇酶不反应,烟曲霉在该表位中仅有两个氨基酸不同。本研究结果表明单克隆抗体1C7在蛋白质免疫印迹应用中检测球毛壳菌和其他粪壳菌纲物种具有潜在用途。