Bacterial Toxin Research Citations

… These assays were conducted using 60.8 ng/ml of PA (List Biological Laboratories, Campbell, CA) and 48.6 ng/ml of LF (List Biological Laboratories) to achieve more than 99% cell lysis at a fixed cell density of 3 10 4 cells/well as determined by toxin potency testing. …

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

Reagents:

… The Anthrax Lethal Factor and Anthrax Protective Antigen were obtained from List Biological Laboratories, INC.

Assay for the screening of the small molecule library against APA and ALF:

An analyzing solution consisting of 10 L of the Small molecule-carrying MRnS
and 2,000 L DI water was prepared. Samples containing different concentrations of free
toxin (APA or ALF, 2 pM to 20 nM, in 1X PBS buffer) were prepared and 2 L of each
sample was added to 200 L of the MRnS analyzing solution (Small molecule-MRnS). A
negative control sample was prepared in the same fashion, adding 2 L fresh 1X PBS
buffer instead of toxin (0 M APA or ALF control sample). Magnetic relaxation
measurements were performed every 15 minutes of incubation at room temperature for 1
hour. …

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

Synthesis of HD5 and Mutant Analogues:

… Recombinant anthrax lethal factor was purchased from List Biological Laboratories, Inc. …

LF Inhibition Kinetics:

The inhibition of LF by various defensins was quantified using an enzymatic kinetic assay (52). Briefly, freshly prepared LF at a final concentration of 1 g/ml (10 nm) was incubated at 37 C for 30 min with a 2-fold dilution series of defensin in 20 mm HEPES buffer containing 1 mm CaCl2 and 0.5% Nonidet P-40, pH 7.2. 20 l of LF substrate (1 mm) was added to each well to a final concentration of 100 m in a total volume of 200 l. The enzyme activity, characterized as a time-dependent absorbance increase at 405 nm due to the release of p-nitroaniline, was monitored at 37 C over a period of 5 min on a 96-well Vmax microplate reader …

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).

Phage display and phage ELISA:

… To measure the activity of phage-displayed scFvs, phage ELISAs were performed in high-binding 96-well plates (Costar) coated with 1 g/ml KJ126 monoclonal antibody (BD Pharmingen; to assess DO11.10 scFv activity), 16 g/ml anthrax protective antigen (List Labs; 14B7), …

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

Materials:

… Protective antigen was acquired from List Biologicals (Campbell, CA), …

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

Recombinant protein reagents:

Recombinant anthrax toxins, rPA, rLF, or rEF (List Biological Laboratories, Campbell, CA) were stored frozen in 1 mg/ml aliquots as recommended by the supplier. …

Protective antigen, lethal factor, and edema factor ELISAs:

Antibody specificity for toxin components was determined using standard ELISAs. High bind plates (Costar 3369) were coated overnight with 1 g/well of rPA, rLF, or rEF. …

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

Author did not specify which List Labs Anthrax Edema Factor was utilized.  List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).

Synthesis and Folding of Defensins:

… Recombinant LF was purchased from List Biological Laboratories, Inc. …

LF Activity Inhibition by l-Ala17-HNP1 Was Less Than Wild-type HNP1:

The enzyme activity of LF in the presence of HNP1 or l-Ala17-HNP1 was assessed using a method adapted from Kaufmann and colleagues (30). Fig. 7 shows that the inhibitory activity of l-Ala17-HNP1 as measured by the concentration that gave 50% inhibition (IC50) differed slightly more than 2-fold compared with HNP1. By contrast, d-Ala17-HNP2 (HNP1 numbering) was equally active compared with HNP2. The d-Ala17 mutation was incorporated into HNP2 instead of HNP1 to conform to the previous study (18), but because HNP1 and HNP2 vary only by the deletion of a single Ala residue at the N terminus, they should be comparable with one another with respect to the internal Gly17 residue. LF inhibition is impaired only by the Gly17 to l-Ala17 mutation but not the Gly17 to d-Ala17 mutation, pointing to a local structural defect as the culprit.

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).

Animal models of anthrax infection:

… Groups of female Hartley guinea-pigs (250-280 g) were immunized by intramuscular injection into the hind leg with 0.2 ml of 50 g of PA (List Biological Laboratories, Inc.), D4 or PDGA-D4 absorbed to 25% aluminum hydroxide (Alhydrogel) on days 0, 14 and 28. …

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

… The B. anthracis proteins, protective antigen (PA), edema factor (EF), and lethal factor (LF) were purchased from List Biological Laboratories, Inc. and required HRP labeling prior to analysis. …

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

Author did not specify which List Labs Anthrax Edema Factor was utilized.  List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).

… Recombinant Bacillus anthracis lethal factor (LF) and Clostridium botulinum neurotoxin light chain A (BoNTALC) were obtained from List Laboratories (Campbell, CA; 169A and 610A, respectively). …

Protease Inhibition Assay:

Protease inhibition assays were performed as described
previously (25), but with modifications. Biotinylated green
fluorescent protein (GFP) protease substrates for LF, BoN-
TALC, and a protease-resistant substrate (pinpointGFP)
were prepared and loaded on streptavidin microspheres as
described previously (14,25,26). Additions to wells were in
sequence as follows: (1) 4 lL protease buffer; (2) 2 lLofa
mixture of 1.5 lM LF and 5 nM BoNTALC in protease
buffer; …

• Product #610A – Botulinum Neurotoxin Type A Light Chain, Recombinant

List Labs Product #169A (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence; Lyophilized) has been discontinued; however, Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence; Liquid) is available for purchase.

• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence