• KSBB Journal
• /
• v.18 no.5
• /
• pp.429-433
• /
• 2003

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) ${\times}$ 18 mm (width) prototype in which 2 mm (width) ${\times}$ 44.34 mm (length) ${\times}$ 100 $\mu\textrm{m}$ (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 ${\mu}\ell$ to 4.4 ${\mu}\ell$. In sum, this study revealed that the mini LOC could be an alternative for a semi-automated and reliable method for LAL test.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.9 no.2
• /
• pp.132-136
• /
• 2004

The LAL (Limulus amebocyte lysate) test for the detection and quantification of endotoxin is based on the gelation reaction between endotoxin and LAL from a blood extract of Limulus polyphemus. The test is labor intensive, requiring dedicated personnel, a relatively long reaction time (approximately 1 h), relatively large volumes of samples and reagents and the detection of the end-point is rather subjective. To solve these problems, a miniaturized LOC (lab-on-a-chip) prototype, 62mm (L) ${\times}$ 18 mm (W), was fabricated using PDMS (polydimethylsiloxane) bonded to glass. Using this prototype, in which 2mm (W) ${\times}$ 44.3mm (L) ${\times}$ 100 $\mu\textrm{m}$ (D) microfluidic channel was constructed, turbidometric and chromogenic assay detection methods were compared, and the chromogenic method was found the most suitable for a small volume assay. In this assay, the kinetic-point method was more accurate than the end-point method. The PDMS chip thickness was found to be minimized to around 2 mm to allow sufficient light transmittance, which necessitated the use of a glass slide bonding for chip rigidity. Due to this miniaturization, the test time was reduced from 1 h to less than 10 min, and the sample volume could be reduced from 100 to ca. 4.4 ${\mu}$L. In summation, this study suggested that the LOC using the LAL test principle could be an alternative as a semi-automated and reliable method for the detection of endotoxin.