title: Development of porcine ficolin-alpha monoclonal and polyclonal antibodies for determining the binding capacity of multiple GlcNAc-binding proteins to bacterial danger components.
abstract: Ficolins are a group of oligomeric defense proteins assembled from collagen-like stalks and fibrinogen-like domains that have common biochemical specificity for N-acetyl-d-glucose amine (GlcNAc) and can function as opsonins. In this report, GlcNAc-binding protein (GBP) purified from porcine nonimmune serum was biochemically characterized as ficolin-α. Ficolin-α was used as an immunogen to generate both rabbit polyclonal and murine monoclonal anti-ficolin-α antibodies, which are not yet commercially available. GBPs have been shown to be present in many animals, including humans; however, their functions are largely unknown. GBPs from chicken, dog, horse, bovine, and human sera were isolated using various chromatography methods. Interestingly, anti-ficolin-α antibody showed cross-reaction with those animal sera GBPs. Furthermore, anti-ficolin-α antibody was reactive with the GlcNAc eluate of Escherichia coli O26-bound and Salmonella-bound porcine serum proteins. Functionally, GBPs and bacteria-reactive pig serum proteins were able to bind with pathogen-associated molecular patterns such as lipopolysaccharides and lipoteichoic acids. Our studies demonstrate that ficolin-α specific antibody was reactive with GBPs from many species as well as bacteria-reactive serum proteins. These proteins may play important roles in innate immunity by sensing danger components that can lead to antibacterial activity.
authors.affiliation: Immunology, Oncology and IMR Discovery Research, Merck Research Laboratories, Boston, MA 021159, USA; Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka 599-853, Japan.
authors.forename: M Abu
authors.initials: MA
authors.lastname: Nahid
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Ross
authors.affiliation: Immunology, Oncology and IMR Discovery Research, Merck Research Laboratories, Boston, MA 021159, USA.
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Umiker
authors.affiliation: Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Huapeng
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka 599-853, Japan.
authors.forename: Sunji
authors.initials: S
authors.lastname: Sugii
authors.affiliation: Center for Advanced Research in Sciences, University of Dhaka, Dhaka 1000, Bangladesh. Electronic address: latiful@du.ac.bd.
authors.forename: Latiful
authors.initials: L
authors.lastname: Bari
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001426:Bacterial Proteins / Q000378:metabolism; D002417:Cattle; D002645:Chickens; D003429:Cross Reactions; D004285:Dogs; D004926:Escherichia coli / Q000378:metabolism; D006736:Horses; D006801:Humans; D037102:Lectins / Q000276:immunology* / Q000302:isolation & purification; D008070:Lipopolysaccharides / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D012486:Salmonella typhimurium / Q000378:metabolism; D013045:Species Specificity; D013552:Swine; D013682:Teichoic Acids / Q000378:metabolism; D000097604:Ficolins
chemical_list: D000117:Acetylglucosamine; D000906:Antibodies; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D037102:Lectins; D008070:Lipopolysaccharides; D013682:Teichoic Acids; D000097604:Ficolins; C009900:lipoteichoic acid
keywords: Cross-reaction; Ficolin; Innate immunity; Monoclonal; Polyclonal antibody