Biotechnology and Bioprocess Engineering:BBE

  • 제9권2호
  • /
  • Pages.132-136
  • /
  • 2004
  • /
  • 1226-8372(pISSN)
  • /
  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Feasibility of On-chip Detection of Endotoxin by LAL Test

  • Lee, Eun-Kyu (Department of Chemical Engineering Hanyang University) ;
  • Suh, Chang-Woo (Department of Chemical Engineering Hanyang University) ;
  • Hwang, Sang-Youn (Department of Chemical Engineering Hanyang University) ;
  • Park, Hyo-Jin (Department of Chemical Engineering Hanyang University) ;
  • Seong, Gi-Hoon (Department of Applied Chemistry Hanyang University) ;
  • Ahn, Yoo-Min (Department of Mechanical Engineering Hanyang University) ;
  • Kim, Yang-Sun (Micro Biochip Center, Hanyang University)
  • 발행 : 2004.03.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

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.

키워드

참고문헌

  1. Clin. Chim. Acta v.307 Microchips, microarrays, biochips and nanochips: Personal laboratories for the 21st century Kricka,L.J. https://doi.org/10.1016/S0009-8981(01)00451-X
  2. Anal. Chem. v.73 Integrated plastic microfluidic devices with ESI-MS for drug screening and residue analysis Yun,J.;P.C.Wang;L.E.Locascio;C.S.Lee https://doi.org/10.1021/ac001474j
  3. Microelec. Eng. v.61;62 PDMS-based microfluidic devices for biomedical applications Fujii,T.
  4. Adv. Drug Delivery Rev. v.55 Lab-on-a-chip for drug development Bernhard,H.W.;L.B.Ron;R.C.Catherine https://doi.org/10.1016/S0169-409X(02)00223-5
  5. Pyrogens: Endotoxins, LAL testing, and Depyrogenation Pearson,F.C.
  6. FEBS Lett. v.129 A new (1→3)-β-d-glucan-mediated coagulation pathway found in Limulus amoebocyte lysate Takashi,M.;T.Shigenori;N.Takanori;I.Sadaaki https://doi.org/10.1016/0014-5793(81)80192-5
  7. Bull. Parenter. Drug. Assoc. v.29 Principle and applications of the Limulus test for pyrogen in parenteral drugs James,F.C.
  8. Trends Biotechnol. v.19 A new era in pyrogen testing Jeak,L.D.;H.Bow https://doi.org/10.1016/S0167-7799(01)01694-8
  9. J. Clin. Microbiol. v.20 Turbidometric method of quantifying serum inhibirion of Limulus amoebocyte lysate Novitsky,T.J.;P.F.Roslansky;G.R.Siber
  10. J. Clin. Microbiol. v.27 Single-step, chromogenic Limulus amoebocyte lysate assay for endotoxin Gene,K.L.;F.R.Priscilla;J.N.Thomas
  11. Dev. Biol. Stand. v.34 Detection of endotoxin in biological products by the Limulus test James,F.C.;M.P.Susan
  12. Korean Pharmacopoeia(7th ed.)
  13. J. Parenter. Sci. Technol. v.46 Viability in the LAL test: Comparison of three kinetic methods for the testing of pharmaceutical products Karen,Z.M.;W.L.Cindy