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KMID : 0380020030180050429
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Korean Journal of Biotechnology and Bioengineering
2003 Volume.18 No. 5 p.429 ~ p.433
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Feasibility Study for a Lab-chip Development for LAL Test
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Hwang Sang-Yeon
Choi Hyo-Jin
Seo Chang-Woo
Ahn Yoo-Min
Kim Yang-Sun
Lee Eun-Kyu
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Abstract
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LAL (Limulus amebocyte lysate) test to detect and quantity endotoxin is based on gellation reaction between endotoxin and LAL from a blood extract of Limulus polyphemus. The test is labor intensive requiring dedicated personnel, takes relatively long reaction time (approximately 1 hr), requires relatively large volume of samples and reagents, and its end-point detection method is rather subjective. To solve these problems, we attempted to develop a miniaturized LOC (lab-on-a-chip) prototype using PDMS and glass. Using the 62 mm (length) 18 mm (width) prototype in which 2 mm (width) 44.34 mm (length) 100 (depth) microfluidic channel was provided, we compared the various detection methods of gellation, turbidometric, and chromogenic assays to find the chromogenic method to be the most suitable for small volume assay. In this assay, kinetic point method was more accurate than end point method. We also found the PDMS chip thickness should be minimized to around 2 mm to allow sufficient light transmittance, which necessitated a glass slide bonding for chip rigidity. Through the miniaturization, the test time was reduced from 1 hr to less than 10 minutes, and the sample volume could be reduced from 100 to 4.4 . In sum, this study revealed that the mini LOC could be an alternative for a semi-automated and reliable method for LAL test.
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KEYWORD
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LAL test, endotoxin, Lab-on-a-chip, PDMS, kinetic chromogenic assay, microfluidic chip
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