Specific immuno capturing of the Staphylococcal superantigen toxic‐shock syndrome toxin‐1 in plasma

Abstract

Toxic-shock syndrome is primarily caused by the Toxic-shock syndrome toxin 1 (TSST-1), which is secreted by the Gram-positive bacterium Staphylococcus aureus. The toxin belongs to a family of superantigens (SAgs) which exhibit several shared biological properties, including the induction of massive cytokine release and V(beta)-specific T-cell proliferation. In this study we explored the possibility to use monoclonal Variable domains of Llama Heavy-chain antibodies (VHH) in the immuno capturing of TSST-1 from plasma. Data is presented that the selected VHHs are highly specific for TSST-1 and can be efficiently produced in large amounts in yeast. In view of affinity chromatography, the VHHs are easily coupled to beads, and are able to deplete TSST-1 from plasma at very low, for example, pathologically relevant, concentrations. When spiked with 4 ng/mL TSST-1 more than 96% of TSST-1 was depleted from pig plasma. These data pave the way to further explore application of high-affinity columns in the specific immuno depletion of SAgs in experimental sepsis models and in sepsis in humans.

Biotechnological applications of recombinant single-domain antibody fragments

“…Multivalency induced by VHH polymerization [97-99] was exploited also to increase the neutralizing capacity of antibodies against the E. coli verotoxin 1 [100], alpha-cobrotoxin [101], and C. difficile toxin A [102], whereas the reduced production costs and the high density at which VHH can be coupled to resin substrates made feasible the preparation of hemofiltration columns for removing toxin shock syndrome toxin 1 from plasma to alleviate staphylococcal-induced sepsis [103,104]. Also an unconventional mouse VH resulting from an incomplete scFv antibody has been isolated and characterized because of its anti-toxin potentialities.…”

“…In a comparative assay with longer antibody constructs and complete IgGs, affinity columns prepared with VHHs enabled higher yields, probably because of the higher density at which they are bound to the matrix [128]. Such VHH-based affinity columns have found their applications for both purifying specific components from heterogeneous material [103,129-131] and for depleting very abundant proteins from samples in which it is necessary to detect the variation of scarcely represented polypeptides. It is the case of the platforms developed for the removal of IgG and human serum albumin from plasma before processing the samples in proteomic assays [132] and of the nanotrap for the purification of GFP-fused proteins from cell homogenates [133,134].…”

Isolation of a Highly Thermal Stable Lama Single Domain Antibody Specific for Staphylococcus aureusEnterotoxin B

“…Previously, sdAbs able to bind small molecules (caffeine and methotrexate), or toxins (botulinum, ricin, cholera, and scorpion), and viruses (rotavirus, HIV, Vaccinia, and Marburg) have been isolated [11,14-20]. Of particular relevance, a sdAb has recently been developed for the related toxin, toxic-shock syndrome toxin 1 (TSST-1),[21] and another for the detection of Staphylococcus aureus [22]. …”

Critical Roles of Clostridium difficile Toxin B Enzymatic Activities in Pathogenesis

“…Because of their small size, V H Hs are often found to have unusual epitope specificities, particularly an improved capability to bind to activesite pockets in enzymes in order to inhibit their functions (46,47). V H Hs have been reported in the treatment of toxin-mediated disease (48,49). V H Hs against the two C. difficile toxins have also been generated which possessed toxin-neutralizing activity (42,50) and potent therapeutic efficacy against fulminant C. difficile infection (51).…”