Production of a monoclonal antibody against a mannose-binding protein of Acanthamoeba culbertsoni and its localization

• Published in: Experimental parasitology 2018-09, Vol.192, p.19-24
• Main Authors: Kang, A-Young, Park, A-Young, Shin, Ho-Joon, Khan, Naveed Ahmed, Maciver, Sutherland K., Jung, Suk-Yul
• Format: Article
• Language: English
• Subjects: Acanthamoeba culbertsoni; Detection; Mannose-binding protein; Monoclonal antibody
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2018.07.009

Amoebae from the genus Acanthamoeba are facultative pathogens of humans and other animals. In humans they most frequently infect the eye causing a sight threatening infection known as Acanthamoeba keratitis (AK), and also cause an often fatal encephalitis (GAE). A mannose-binding protein (MBP) has been identified as being important for Acanthamoeba infection especially in AK. This lectin has previously been characterized from Acanthamoeba castellanii as consisting of multiple 130 kDa subunits. MBP expression correlates with pathogenic potential and is expressed in a number of Acanthamoeba species. Here we report the purification of a similar lectin from Acanthamoeba culbertsoni and the production of a monoclonal antibody to it. The A. culbertsoni MBP was isolated by affinity chromatography using α-D-mannose agarose and has an apparent molecular weight of 83 kDa. The monoclonal antibody is an IgM that is useful in both western blots and immunofluorescence. We expect that this antibody will be useful in the study of the pathology of A. culbertsoni and in its identification in clinical samples. [Display omitted] •A monoclonal antibody to A. culbertsoni MBP was produced. It was used in immunocytochemistry to localize MBP and may be useful in diagnosis of GAE.•The monoclonal antibody was produced by the cell-fusion technique and characterized.•The antibody will be useful in the study of the pathology of A. culbertsoni and in its identification in clinical samples.