title: Domain-specific monoclonal antibodies produced against human PGRN.
abstract: Progranulin (PGRN) encodes a 68.5-kDa secreted growth factor that is composed of seven and a half tandem repeats of a 12-cysteine granulin motif. PGRN is expressed in many tissues and has a role in mediating development, wound repair, inflammation, and tumorigenesis. Mutations leading to a loss of function in PGRN are the most common cause of familial frontotemporal lobar degeneration with TDP-43-positive inclusions (FTLD-TDP). In this study, we established hybridoma cell lines producing antibodies against human PGRN. Murine monoclonal antibodies (MAbs) against human PGRN were generated by using purified eukaryotic recombinant PGRN-6His fusion protein as immunogen. A panel of seven monoclonal antibodies was obtained after the preliminary screening by indirect enzyme-linked immunosorbent assay (ELISA), the data of which was confirmed by Western blot analysis and immunocytochemistry. By using constructs expressing a series of C- and N-terminal truncations, and single domains of PGRN, the particular domains recognized by MAbs were also identified. Domain-specific anti-PGRN MAbs will be an essential tool for investigating the role of PGRN in normal physiological or pathological conditions.
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, PR China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Chai
authors.forename: Qinwen
authors.initials: Q
authors.lastname: Mao
authors.forename: Shihai
authors.initials: S
authors.lastname: Liu
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000918:Antibody Specificity; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D000077153:Progranulins; D011993:Recombinant Fusion Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; C492314:GRN protein, human; D036341:Intercellular Signaling Peptides and Proteins; D000077153:Progranulins; D011993:Recombinant Fusion Proteins