Bacterial Toxin Research Citations

In vitro BoNT LC metalloprotease activity:

Determination of BoNT/A LC percent inhibition by SMNPIs was performed as previously described.  For BoNT/B LC percent inhibition, the HTS-assay utilizes a commercially available fluorogenic substrate from List Biological Laboratories (Campbell, CA). Briefly, a multichannel pipette was used to add fluorescent substrate (final concentration 20 M) and small molecules (final concentration 20 M) to a 96 well microplate. The reactions were initiated by adding recombinant BoNT/B LC (final concentration 40 nM) to each well. All reactions were conducted in a buffered solution consisting of 40 mM Hepes pH 7.2, 1 mM DTT and 100 M ZnCl2. The change in fluorescence intensity over time was continuously monitored in each well using a 96 well plate fluorimeter (Saphire 2, Tecan, Mnnedorf, Switzerland). The assays were run at 37 C, quenched by the addition of TFA, and analyzed using reverse-phase HPLC. Potential SMNPIs of the BoNT B LC were identified by comparing the reaction velocities (change in fluorescence intensity over time) of samples assayed in the presence of compounds versus  control samples.

Author did not specify which List Biological Laboratories product was utilized.  Refer to the follwing List Labs link for Botulinum products available:  https://listlabs.com/product-information/botulinum-toxins/

Cell treatments:

For Western blot experiments and luciferase assays the following treatment regimen was used. Six hours prior to harvesting, cells were treated with 10 mg/ml medium of Escherichia coliderived LPS (E. coli 055:B5; List Biological Laboratories, Campbell, CA), …

NOTE:  At the time this paper was written, Product #203 (5mg – LPS from Escherichia coli O55:B5) was utilized.

Product #203 (5 mg – LPS from Escherichia coli O55:B5) is no longer sold.

Product #203A (2.5 mg – LPS from Escherichia coli O55:B5) is available for purchase.

• Product #203A – LPS from Escherichia coli O55:B5

rPA Delivery in Rats:

Recombinant protective antigen from Bacillus anthracis (rPA) was obtained from List Biologicals (Campbell, CA). The MSA containing rPA was fabricated using a casting and drying fabrication method. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Anti-PA and anti-LF IgG and IgA antibody titers:

Anti-PA and anti-LF immunoglobulin G (IgG) titers in mouse serum samples were determined by enzyme-linked immunosorbent assay (ELISA) using purified PA or LF protein (List Biological Laboratories, Campbell, CA) as the capture antigen. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Preparation of bacteria culture filtrate:

… A standard curve was generated using serially diluted commercial toxin A or toxin B products with known concentrations (List Biological Lab, 152C and 155B, respectively). Standard curves were plotted as the standard toxin A or toxin B protein concentration (ng/ml) against the corresponding mean OD value of replicates. …

• Product #152C – Toxin A from Clostridium difficile
• Product #155B – Toxin B from Clostridium difficile

Administration of BCG and CT:

The M. bovis BCG Tokyo 172 strain (Japan BCG Laboratory, Tokyo, Japan) was prepared as… CT and CTB were purchased from List Biological Lab, Campbell, CA. Groups of mice (6 mice/group) were given 50 µl of saline containing 500 µg of BCG and 1 µg of CT, 500 µg of BCG, 1µg of CT, or saline (controls) intranasally at 0 h, and samples were collected at 24 h. The intranasal doses of BCG and CT were based on previous studies…

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Sensitization:

Mice were orally sensitized to peanut by intragastric administration of 10mg ground peanut (prepared from shelled peanuts) in 0.2m sodium bicarbonate with 10µg of CT (List Biologicals, Campbell, CA) once a week for 6 weeks.

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Cholera toxin (List Biological Labs, Campbell, CA) was eliminated from KCM for canine and rat epithelial cell culture, but added at a concentration of 1 nm for human epidermal keratinocyte culture.

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Murine peanut allergy model:

… The mice were orally sensitized with 500 g of WPE along with 10 µg of cholera toxin (List Biological Laboratories, Campbell, Calif) once a week for 4 weeks. …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Peanut plus CT administration:

One milligram of peanut (Kraft, Northfield, Ill) with 10 µg of CT (List Biological Laboratories, Campbell, Calif) in PBS was gavaged intragastrically by using 12-gauge, …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae