Anti-HA antibody does not detect hyaluronan

Abstract

It is generally known that hyaluronic acid (HA) is a biocompatible and biodegradable glycosaminoglycan distributed widely throughout epithelial, connective and neural tissues. HA is one of the chief components of the extracellular matrix. Lack of immunogenicity is one of the biggest advantages of the therapeutic use of HA, but it also prevents the production of specific anti-HA antibodies. Contrary to this, there are still several studies performing HA detection by immunohistochemical or immunohistofluorescent method using an anti-HA antibody. Therefore, this short study discusses whether the anti-HA antibody is specific for HA. To verify the specificity of the HA staining the hyaluronidase treatment of histological samples was performed and the ability of anti-HA antibody and biotinylated HA binding protein (bHABP)-based probe to bind to their targets was evaluated. Additionally, the competitive binding assay with short HA oligosaccharides and subsequent histological staining was performed. Both assays showed that the anti-HA antibody is not sufficiently specific for HA and that the bHABP probe is a reliable method for HA detection in histological samples. The conclusion made by previous investigators based on using HA antibodies should be reevaluated and future use of anti-HA antibody should be avoided.

Keywords: IFC; IHC; anti-HA antibody; hyaluronan; hyaluronic acid binding protein; hyaluronidase.