Conventional Test Method:

Limulus Amebocyte Lysate (LAL) Test a.k.a. Bacterial Endotoxin Test

Alternative Test Method:

Name & Description:

Recombinant Factor C (rFC) Bacterial Endotoxin Test

Recombinant Factor C (rFC) is a coagulation enzyme precursor (proenzyme or zymogen) found in LAL which is catalytically activated by endotoxin. rFC is incubated with test substance and other substrates which aid in endotoxin detection or quantification. Detection techniques include a chromogenic (colour-producing) assay, a fluorogenic (fluorescence-producing) assay and ELISA with chromogenic substrate. Endotoxin presence is measured by the amount of colour or fluorescence produced.

Validation Status:

US: Currently being compared to the Bacterial Endotoxin Test for inclusion in the USP

Regulatory Status:

No information

Effect or Potential Effect on Animal Use:

Replacement

References:

• Ding J.L. & Ho B. (2001) A new era in pyrogen testing. Trends in Biotechnology 19: 277-281.
• Tan N.S., Ho B. & Ding J.L. (2000) High-affinity LPS binding domain(s) in recombinant factor C of a horseshoe crab neutralizes LPS-induced lethality. FASEB 14: 859-870.
• Ding J.L. & Ho B. (2010) Endotoxin detection- from limulus amebocyte lysate to recombinant factor C. Subcellular Biochemistry 53: 187-208.

Information Last Reviewed:

May 2012