Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Video

Updated: Mar 11, 2026

Air-sampled Filter Analysis for Endotoxins and DNA Content

Air-sampled Filter Analysis for Endotoxins and DNA Content

Published on: March 7, 2016

Genetic engineering approach to develop next-generation reagents for endotoxin quantification.

Hikaru Mizumura¹, Norihiko Ogura¹, Jun Aketagawa²

¹1 LAL Research and Development Department, Seikagaku Corporation, Tokyo, Japan.

Innate Immunity

|December 4, 2016

Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Nodular Scleroderma in a Patient With Anti-RNA Polymerase III-Antibody Positive Systemic Sclerosis: A Case Report and Literature Review.

The Journal of dermatology·2026

Same author

Hydrogen-generating silicon-based agent is effective in a mouse model of ovalbumin-induced allergic bronchial asthma.

Biomedical journal·2026

Same author

Delayed-onset parkinsonism possibly associated with elranatamab treatment for relapsed/refractory multiple myeloma: a case report of a poorly characterized neurological event.

Annals of hematology·2026

Same author

Ectomesenchyme contributes to epidermal stem cell formation through mesenchymal-to-epithelial transition.

Nature communications·2026

Same author

Structural basis of nucleosome remodeling by Cockayne syndrome B homologue Komagataella phaffii Rad26.

Nature communications·2026

Same author

Comparative earthquake sensitivity of straight and triply helical fibers in surface distributed acoustic sensing.

Scientific reports·2026

This study developed sustainable recombinant cascade reagents (RCRs) for bacterial endotoxin testing, replacing horseshoe crab-derived lysate. These RCRs show enhanced sensitivity and reduced interference, paving the way for eco-friendly diagnostics.

Area of Science:

Biotechnology
Biochemistry
Analytical Chemistry

Background:

The bacterial endotoxin test (BET) traditionally relies on amebocyte lysate (LAL) derived from horseshoe crabs.
Horseshoe crab populations are facing pressure, necessitating sustainable alternatives for LAL reagents.

Purpose of the Study:

To develop sustainable, recombinant raw materials for lysate reagents.
To engineer recombinant cascade reagents (RCRs) that mimic the natural LAL reaction cascade.
To evaluate the performance of RCRs as an alternative to traditional LAL reagents.

Main Methods:

Genetic engineering was used to prepare three recombinant protease zymogens: factor C (from mammalian cells), factor B, and proclotting enzyme (from insect cells).
Recombinant cascade reagents (RCRs) were constructed to replicate the amebocyte lysate reaction cascade.

Keywords:

Amoebocyte lysate reagent endotoxin horseshoe crab mammalian cell recombinant cascade reagent

More Related Videos

Production of E. coli-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Production of E. coli-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Published on: August 21, 2019

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

Published on: June 24, 2019

Related Experiment Videos

Last Updated: Mar 11, 2026

Air-sampled Filter Analysis for Endotoxins and DNA Content

Air-sampled Filter Analysis for Endotoxins and DNA Content

Published on: March 7, 2016

Production of E. coli-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Production of E. coli-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation

Published on: August 21, 2019

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients

Published on: June 24, 2019

The protease activity, signal amplification efficiency, and endotoxin sensitivity of RCRs were compared to natural LAL reagents.

Main Results:

RCRs demonstrated efficient signal amplification, with protease activity significantly higher than recombinant factor C alone.
RCRs containing mammalian cell-derived factor C showed reduced interference from injectable drug components compared to insect cell-derived factor C.
RCRs with mammalian cell-derived recombinant factor C exhibited a steeper standard curve slope, indicating greater endotoxin sensitivity than natural LAL reagents.

Conclusions:

Recombinant reagents can be produced without relying on natural resources like horseshoe crabs.
Mammalian cell-derived recombinant factor C-based RCRs offer improved sensitivity and reduced interference for endotoxin testing.
This study supports the future production of sustainable, recombinant reagents for bacterial endotoxin detection.