title: A quantitative ELISA assay for the fragile x mental retardation 1 protein.
abstract: Non-coding (CGG-repeat) expansions in the fragile X mental retardation 1 (FMR1) gene result in a spectrum of disorders involving altered neurodevelopment (fragile X syndrome), neurodegeneration (late-onset fragile X-associated tremor/ataxia syndrome), or primary ovarian insufficiency. While reliable and quantitative assays for the number of CGG repeats and FMR1 mRNA levels are now available, there has been no scalable, quantitative assay for the FMR1 protein (FMRP) in non-transformed cells. Using a combination of avian and murine antibodies to FMRP, we developed a sensitive and highly specific sandwich enzyme-linked immunosorbent assay (ELISA) for FMRP in peripheral blood lymphocytes. This ELISA method is capable of quantifying FMRP levels throughout the biologically relevant range of protein concentrations and is specific for the intact FMRP protein. Moreover, the ELISA is well-suited for replicate protein determinations across serial dilutions in non-transformed cells and is readily scalable for large sample numbers. The FMRP ELISA is potentially a powerful tool in expanding our understanding of the relationship between FMRP levels and the various FMR1-associated clinical phenotypes.
authors.affiliation: Department of Biochemistry and Molecular Medicine, UC Davis, School of Medicine, Davis, CA 95616, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Iwahashi
authors.forename: Flora
authors.initials: F
authors.lastname: Tassone
authors.forename: Randi J
authors.initials: RJ
authors.lastname: Hagerman
authors.forename: Dag
authors.initials: D
authors.lastname: Yasui
authors.forename: George
authors.initials: G
authors.lastname: Parrott
authors.forename: Danh
authors.initials: D
authors.lastname: Nguyen
authors.forename: Greg
authors.initials: G
authors.lastname: Mayeur
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hagerman
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000097:blood* / Q000235:genetics; D005600:Fragile X Syndrome / Q000097:blood / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D002352:Carrier Proteins; D051860:Fragile X Messenger Ribonucleoprotein 1; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins