Transforming growth factor type beta in normal human urine.
TGF beta has been identified in normal human urine specimens from five individuals studied for five consecutive days. The peptide was extracted from urine using Sepralyte C1 beads. Detectable levels of [125I]TGF beta competing activity as measured by radioreceptor assay was found in about half of the specimens studied. The protein isolated from urine using C1 Sepralyte beads was further purified using Biogel P-60 column chromatography. Fractions were tested for TGF beta and EGF competing activity using radioreceptor assays. TGF beta and EGF extracted from urine are clearly separated by column chromatography. Two distinct EGF peaks and a single TGF beta peak were observed. Fractions having [125I]TGF beta competing activity were pooled and further purified using reverse-phase HPLC. HPLC fractions having [125I]TGF beta competing activity were tested for bioactivity using a soft agar assay. The fractions were capable of stimulating soft agar growth of AKR-2B (clone 84A) cells and cross reacted with a TGF beta antibody in a radioimmunoassay. The presence of TGF beta in normal human urine was also demonstrated by immunoblotting. These results also suggest that C1 bead extraction of urine specimens can be used as a rapid first step in purification of TGF beta.
Ranganathan
G
G
Department of Laboratory Medicine and Surgical Pathology, Mayo Clinic, Rochester, Minnesota.
Lyons
R
R
Jiang
N S
NS
Moses
H
H
D001681:Biological Assay; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002940:Circadian Rhythm; D006801:Humans; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D010455:Peptides / Q000652:urine*; D011869:Radioligand Assay; D015291:Transforming Growth Factors
D010455:Peptides; D015291:Transforming Growth Factors