Comparative urine protein phenotyping using mass spectrometric immunoassay.
Reported here, human urine samples were analyzed for beta-2-microglobulin (beta2m), transthyretin (TTR), cystatin C, urine protein 1 (UP1), retinol binding protein (RBP), albumin, transferrin, and human neutrophil defensin peptides (HNP) using mass spectrometric immunoassay (MSIA). MSIA is a unique analytical technique, which allows for the generation of distinct protein profiles of specific target proteins from each subject, which may be subsequently used in comparative protein expression profiling between all subjects. Comparative profiling allows for the rapid identification of variations within individual protein expression profiles. Although the majority of analyses performed in this study revealed homology between study participants, roughly one-quarter showed variation in the protein profiles. Some of these observed variants included a point mutation in TTR, absence of wild-type RBP, monomeric forms UP1, a novel beta2m glycated end product and altered HNP ratios. MSIA has been previously used in the analysis of blood proteins, but this study shows how MSIA easily transitions to the analysis, of urine samples. This study displays how qualitative urine protein differentiation is readily achievable with MSIA and is useful in identifying proteomic differences between subjects that might be otherwise overlooked with other analytical techniques due to complexity of the resulting data or insufficient sensitivity.
Kiernan
Urban A
UA
Intrinsic Bioprobes, Inc., 625 South Smith Road, Suite 22, Tempe, Arizona 85281, USA.
Tubbs
Kemmons A
KA
Nedelkov
Dobrin
D
Niederkofler
Eric E
EE
McConnell
Elizabeth
E
Nelson
Randall W
RW
D004252:DNA Mutational Analysis; D020869:Gene Expression Profiling / Q000379:methods; D014644:Genetic Variation; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D040901:Proteomics / Q000379:methods; D012680:Sensitivity and Specificity; D014556:Urine / Q000737:chemistry*
D011506:Proteins