Effects of Boric Acid and Storage Temperature on the Analysis of Microalbumin Using Aptasensor-Based Fluorescent Detection

硼酸和储存温度对适体传感器荧光检测微量白蛋白的影响

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作者:Chalermwoot Sompark, Wireeya Chawjiraphan, Manatsaphon Sukmak, Ubon Cha'on, Sirirat Anutrakulchai, Prapasiri Pongprayoon, Thitirat Putnin, Dechnarong Pimalai, Visarute Pinrod, Deanpen Japrung

Abstract

The instability of human serum albumin (HSA) in urine samples makes fresh urine a requirement for microalbumin analyses using immunoturbidimetry. Here, we determined the ability of an aptasensor-based fluorescent platform to detect microalbumin in old, boric acid-preserved urine samples. Our results show that the cleavage site of protease enzymes on urine albumin protein differed from the binding position of the aptamer on HSA protein, suggesting the aptasensor may be effective for albumin detection in non-fresh urine. Furthermore, the addition of boric acid in urine samples over a short term (at ambient temperature (Ta) and 4 °C), long term (-20 and -80 °C), and following freeze-thawing (1-3 cycles) did not significantly affect albumin stability, as analyzed using the aptasensor. Therefore, boric acid stabilized has in urine stored over a short- and long-term. Thus, the aptasensor developed by us is applicable for HSA detection in boric acid-preserved urine that has been stored for 7-d at Ta and 4 °C, and in the long-term at -80 °C.

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