Endotoxin detection

• Endotoxin detection assays
• Instrumenter & Software
• Accessory items
• Endotoxin detection services

Endotoxin detection

Endotoxin detection assays

Endotoxins, a type of pyrogen, are natural compounds found in the outer cell membrane of Gram-negative bacteria and impacts over 30 biological activities. Endotoxins can lead to cell death by initiating complement activation. 

Endotoxin detection tests are performed on raw and in-process materials, and for the final release of products in the pharmaceutical and medical device industries.Using extensive experience with endotoxin detection and its regulatory requirements, Lonza has been able to deliver innovative and time-saving tools for both the pharmaceutical and research markets.
 

Limulus Amebocyte Lysate (LAL) tests

The Limulus Amebocyte Lysate (LAL) test was commercially introduced in the 1970s. LAL is derived from the blood cells, or amebocytes, of the horseshoe crab, Limulus polyphemus. Frederick Bang and Jack Levin observed that blood cells from horseshoe crabs were found to clot in the presence of endotoxin, and this technology was used in the development of endotoxin detection assays.

Lonza offer several types LAL tests, both qualitative and quantitative: 

PyrogentTM Gel Clot LAL Assay (qualitative assay):
• Visual inspection of gel formulation
• Maximum sensitivity: 0.03 EU/ml
• Instrument required: dry heat block or water bath

Kinetic-QCLTM Kinetic Chromogenic LAL Assay (quantitative assay):
• Kinetic measurement of color development
• Maximum sensitivity: 0.005 EU/ml
• Our most sensitive LAL-based method
• Less sensitive to product inhibition
• Ideal for biological products such as vaccines and antibiotics
• Instrument required: absorbance reader

PyrogentTM-5000 Kinetic Turbidimetric LAL Assay (quantitative assay):
• Kinetic measurement of turbidity development
• Maximum sensitivity: 0.01 EU/ml
• Cost-effective method for water and large volume parenterals
• Instrument required: absorbance reader

QCL-1000TM Endpoint Chromogenic LAL Assay (quantitative assay):
• Endpoint measurement of color development
• Maximum sensitivity: 0.01 EU/ml
• Results in 16 minutes
• Instrument required: spectrophotometer or absorbance reader, dry heat block
 

Endotoxin detection assays
Endotoxin detection assays
Recombinant Factor C Endpoint Fluorescent Assay
Cat. nos. 50-658U (192 tests) and 50-658NV (2880 tests)

Today, an animal-free alternative to LAL is also available; the PyroGene Recombinant Factor C Assay. It is based on a recombinantly produced form of Factor C (rFC), the first component in the horseshoe crab clotting cascade. It is activated by endotoxin binding. The active moiety created then acts to cleave a synthetic substrate, which results in the release of a fluorophore. The reaction is run in a 96-well microplate and measured at time zero and after a one-hour incubatyion in a fluorescence microplate reader using excitation/emission wavelenghts of 380/440 nm. 

• Endpoint measurement of fluorescence
• Sensitivity range from 0.005 to 50 EU/ml
• Higher endotoxin sensitivity
• Elimination of false positive glucan reactions
• Less lot-to-lot variability
• Animal-free source and security of supply
• Instrument required: incubating fluoresence reader

For detailed product information about the different detection methods/assay available, contact us, or read more here.

Instruments & Software

Lonza is the industry leader in innovations for endotoxin detection products, services and software. Using their extensive experience with endotoxin detection and its regulatory requirements, they developed an integrated system to support quantitative endotoxin detection. Each system component has been validated and can be verified.

Two plate readers have been validated as part of their quantitative endotoxin detection systems. These 96-well plate readers feature sample compartments with uniform temperature from well to well, and both readers seamlessly interface with their WinKQCL software.

Instruments & Software
Instruments & Software
ELx808TM incubating absorbance plate reader
Cat. no. 25-315S

The ELx808TM Incubating Absorbance Microplate Reader has been validated as part of Lonza's quantitative endotoxin detection system interfacing with Lonza's WinKQCLTM Endotoxin Detection and Analysis Software. This 96-well microplate reader features 4 insulated incubating zones, providing well-to-well temperature uniformity available on the market. Utilizing interference filters instead of monochromators, the reader is optimized for the LAL-based endotoxin tests and comes pre-configured with the following interference filters: 340nm, 405nm, 450nm, 490nm, and 630nm. The ELx808TM Reader seamlessly interfaces with Lonza's  WinKQCLTM Endotoxin Detection and Analysis Software. 

For the kinetic and endpoint LAL assays, the ELx808TM Reader is highly reliable and easy to use. It features fully-automated kinetic reads of the 96-well plate via the WinKQCLTM Software. This reader can also be used as a standard spectrophotometer and results can be read from the QCL-1000TM Endpoint Chromogenic LAL Assay and ELISA Assays. 

ELx808TM Absorbance Reader Specifications
Wavelength range 340 to 900 nm
Filters supplied 340, 405, 450, 490, and 630 nm
Absorbance range 0.000 to 4.000 OD @ 400 to 900 nm
0.000 to 3.000 OD @ 340 to 400 nm
Temperature control  4°C above ambient to 50°C
Read method Kinetic or endpoint under WinKQCLTM control
Light Source Tungsten halogen bulb
Dimensions 40.6 cm x 39.4 cm x 22.2 cm
Weight 15.9 kg

 

PyroWaveTM XM Fluorescence Reader
Cat. no. 25-345S

The PyroWaveTM XM Fluorescence Reader is a part of the quantitative endotoxin detection system that supports the PyrogeneTM rFC Endpoint Fluorescent Assay. This reader replaces the Lonza Flx800TM LBS reader, bringing in a new generation in fluorescence technology to users of the PyrogeneTM Assay. Optimized specifically for Lonza's PyrogeneTM, this reader brings new and improved technology to the laboratory. The PyroWaveTM XM reader offers numerous enhancements in incubation, optics, automation capability, and overall reader maintenanceand robustness. Controlled by Lonza's WinKQCLTM Endotoxin and Analysis Software, version 5.3 and higher, Lonza delivers a high performance and easy to use system for users interested in an alternative to the horseshoe crab-based endotoxin detection test methods.

Technical specifications
Read capabilities Fluorescence, luminescence**, TRF**, FP**
Read position Top read
Light source Xenon flash
Detection High performance multiplier tube
Fluorescence sensitivity Fluorescein @ 1pM/well in a 96 well plate
Wavelength range 200 – 850nm**
Temperature control ±0.2°C at 37°C
Power 100-240 volts AC 50/60 Hz
Dimensions 39.1 cm W x 47.2 cm D x 32.8 cm H
 **Additional Filter cubes and filters must be purchased for additional wavelengths and read capabilities

One easy-to-swap filter cube is included, with the following filter configuration:

Assay Type Excitation Filter (nm) Emission Filter (nm) Dichroic Filter (nm)
Pyrogene 380/20 440/30 400
Fluorscein 485/20 528/20 510
WinKQCL endotoxin detection and analysis software

Quantitative methods such as the Kinetic-QCL assay and PyroGene recombinant factor C assay generate significant amounts of raw data that require careful analysis before reporting can take place. The WinKQCL software offers a fully integrated solution for your quantitative endotoxin detection testing, data management, and reporting needs. 

WinKQCL software meets 21 CFR Part 11 technical requirements for electronic records, signatures, audit trails and database archiving. Reader validation tests can be run from the software and are stored in the same database. The built-in backup and maintenance scheduler makes it easy to maintain the system. 
 

Contact us for more information or a quotation. 

Accessory Items

In addition to the endotoxin detection assay kits, instruments and software, Lonza offers all of the accessory items necessary to run endotoxin detection assays.

Many of the items have been tested with the Kinetic-QCL™ Kinetic Chromogenic LAL Assay to help ensure their compatibility with our endotoxin detection methods. They also offer products such as Endotoxin Challenge Vials™ to help customers with oven depyrogenation validations.

Accessory products offered:

Pyrogen-free test tubes (certified to contain less that 0.005 EU/ml endotoxin) recommended for use as reaction tubes in gel clot assays and for dilution of endotoxin standards and test samples. Two sizes available

Sample containers intended for transporting product samples for endotoxin analysis or sample storage.

LAL reagent grade multi-well plates, certified to contain less than 0.0005 EU/well endotoxin and to be free from inhibition

Pyrogen-free pipette tips, certified to contain <0.001 EU/ml or <0.005 EU/ml endotoxin

LAL reagent reservoirs (#190035: 10 reservoirs/pack) for use with multi-channel pipettors when adding reagents to a 96-well plate

Accessory Items
Accessory Items

LAL reagent water is recommended for reconstituting LAL reagents, as well as for the dilution of control standard endotoxin and test samples for endotoxin testing. Certified to contain less than 0.005 EU/ml endotoxin, and certified for positive product control (PPC) recovery within 75 to 150 %. 

beta-G-blocker (#N190: Glucan blocker, 5 ml/vial). Functionality tested to demonstrate a reduction of enhancement caused by beta-D-glucans

PYROSPERSE dispersing agent (#N188: 5 ml/vial). PYROSPERSE dispersing agent helps eliminate endotoxin binding or masking in some samples - solving problems of inhibitory behaviour. 

MgCl2 (#S50-641: MgCl2 10 mM solution, 30 ml/vial). MgCl2 can be used as the sample diluent when attempting to overcome inhibitory chelation effects. 

Tris buffer (#S50-642: Tris buffer 50 mM, 30 ml/vial). Tris buffer can be used in place of water as the sample diluent for highly acidic or basic samples (for endotoxin testing, samples and sample dilutions should be between pH 6-8 after lysate addition). 

Endotoxin and endotoxin challenge vials

Contact us for more details, please see here.

Endotoxin testing services

Lonza's Endotoxin Testing Service combines their expertise with full service reporting, offering both research and pharmaceutical customers’ routine and customized endotoxin testing.

Their facilities provide expertise in gel clot, kinetic chromogenic and kinetic turbidimetric LAL methods, as well as the rFC endpoint fluorescent assay. 

Lonza’s Endotoxin Detection Testing laboratories are located in their cGMP-certified manufacturing plants in Walkersville, MD USA and in Verviers, Belgium.

Endotoxin testing services

All pharmaceutical grade endotoxin testing is performed in accordance with current regulatory documents such as the U.S. and European Pharmacopeia (USP and EP) Bacterial Endotoxins Test (BET), the U.S. Food and Drug Administration (USFDA) Guidance for Industry and the USP monograph for medical devide testing. 

Whether you are from a university, research laboratory or a Quality Control laboratory in a major pharmaceutical company, Lonza has the capabilities you need for reliable, accurate and confidential results

Contact us for more information.