A quantitative ELISA assay for the fragile x mental retardation 1 protein.
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.
Iwahashi
Christine
C
Department of Biochemistry and Molecular Medicine, UC Davis, School of Medicine, Davis, CA 95616, USA.
Tassone
Flora
F
Hagerman
Randi J
RJ
Yasui
Dag
D
Parrott
George
G
Nguyen
Danh
D
Mayeur
Greg
G
Hagerman
Paul J
PJ
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
D002352:Carrier Proteins; D051860:Fragile X Messenger Ribonucleoprotein 1; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins