[Various methods for purification of urine Tamm-Horsfall protein].
The measurements of serum, urine Tamm-Horsfall protein (THP) and its antibody are essential to the early diagnosis of tubulointerstitial diseases and acute rejection after renal transplantation. The purification of THP proves to be the key procedure for the production of diagnostic kit. Once, twice and three times salt precipitation with 0.58 mol/L NaCl and chromatography with Sephadax G-200 were adopted to purify urine THP. Enzyme immunoassay was used to detect the antigenicity of the products, SDS-PAGE to determine the molecular weight (MW) and the electrophoretic purity (EP), Beckman DU-7 model spectrophotometer to scan the ultraviolet absorption spectrum. Evaluation of the four methods was based on the purity and the antigenicity of the produced THP, the output, the complexity of procedure, and the cost of required materials. Results show that the MW of THP is 93.0 kd; the ultraviolet absorption maximum is at lambda = 278 nm; the outputs are 7.5, 5.5, 3.8, and 0.4 mg/L; the concentrations of THP with antigenicity are 65%, 75%, 98% and 100%, respectively, and the EPs of three times salt precipitation and chromatography are better. It is concluded that three times salt precipitation should be recommended because of its excellence in purity and antigenicity of the produced THP, the simplicity of manipulation, and the cheapness of the materials needed.
Du
Y
Y
Xu
G
G
Zhong
L
L
Liu
X
X
D006801:Humans; D008722:Methods; D009088:Mucoproteins / Q000302:isolation & purification* / Q000652:urine; D013056:Spectrophotometry, Ultraviolet; D058949:Uromodulin
D009088:Mucoproteins; C548612:UMOD protein, human; D058949:Uromodulin