Immunoaffinity resin for purification of urinary leukotriene E4.
Urinary leukotriene E4 (LTE4) has been used as an index of total leukotriene synthesis. A wide variety of methods have been applied to measure LTE4 which has made direct comparison of urinary levels reported by different laboratories difficult. A new peptidoleukotriene immunoaffinity resin was utilized for urinary LTE4 purification in a method that is easy and inexpensive, utilizing commercially available reagents. This method is described and compared to other methods. LTE4 (50-250 pg/mL) added to a urine extract was quantitatively recovered using the immunoaffinity resin. Similarly, LTE4 (50-400 pg/mL) added to urine was recovered between 63 and 76%. The coefficient of variation of samples purified and quantified on the same or on different days ranged from 8-10%. There was a strong correlation (r2 = 0.95) between LTE4 concentrations determined after immunofiltration and immunoaffinity purification. Although there was a good correlation between urinary LTE4 levels measured without purification compared to after immunoaffinity purification, the high y-intercept of 179 indicates the presence of interfering substances in unpurified urine. Urinary LTE4 in normal healthy adults was 80 +/- 7 pg/mg creatinine, similar to that previously reported following HPLC or immunofiltration purification. Urinary LTE4 was also measured in healthy children (age 3-12) and found to be 103 +/- 9.
Westcott
J Y
JY
National Jewish Medical and Research Center, Denver, CO 80206, USA. WestcottJ@njc.org
Maxey
K M
KM
MacDonald
J
J
Wenzel
S E
SE
D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D002846:Chromatography, Affinity / Q000379:methods*; D006433:Hemocyanins / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D007164:Immunosorbents; D017999:Leukotriene E4 / Q000276:immunology* / Q000652:urine*; D008875:Middle Aged; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D007164:Immunosorbents; D017999:Leukotriene E4; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin