Engineered Lactic Acid Bacterium Lactococcus lactis Capable of Binding Antibodies and Tumor Necrosis Factor Alpha

Abstract

We have optimized the display of the B domain of staphylococcal protein A on the surface of Lactococcus lactis. The maximum binding capacity was estimated at 0.146 μg of antibody per 108 cells and was sustained at 86% after treatment with simulated gastric juice. A tumor necrosis factor alpha (TNF-α)-binding affibody was also displayed and bound TNF-α, which could be useful in the treatment of inflammatory bowel disease.

Improvement of LysM-Mediated Surface Display of Designed Ankyrin Repeat Proteins (DARPins) in Recombinant and Nonrecombinant Strains of Lactococcus lactis and Lactobacillus Species

“…The expression levels of the two DARPin fusion proteins were the same, and we attribute the stronger IgG binding with I_07-cA to the more complex folding of the larger DARPin, DARPin I_19 (containing 5 ankyrin repeats), than of DARPin I_07 (containing 4 ankyrin repeats). The display of human IgG-binding DARPins on the surface of L. lactis adds another class of LAB-displayed binders to the existing pool, which includes Affibody molecules, Nanobodies, and single-chain variable fragments (11,35,36).…”

“…We previously demonstrated LysM-mediated surface display of Affibody molecules (11). Here we introduced designed ankyrin repeat proteins (DARPins) that bind the Fc domain of human IgGs (27) as model proteins for testing LysM-mediated surface binding.…”

“…Several approaches to the display of heterologous proteins on the surfaces of LAB have been established, including the use of LPXTG-type domains (5), lysine motif (LysM) domains (6), surface layer proteins (7), and lipoprotein anchors (8). Display of heterologous proteins on the surfaces of LAB has been exploited for the preparation of mucosal vaccines (9,10), for the delivery of binding molecules to the gastrointestinal tract (11), for the assembly of macromolecular enzyme complexes (12), and for bacterial immobilization (13).…”

“…It comprises an enzymatic domain at the N terminus and a peptidoglycan-binding domain at the C terminus that comprises 3 LysM repeats (14,16,17). cA has been used as a fusion partner for noncovalent attachment of heterologous proteins to the surfaces of LAB, e.g., attachment of malaria antigen to Lactobacillus sakei and Lactobacillus casei (14), attachment of ␣-amylase to Lactobacillus plantarum and Lactobacillus casei (18), attachment of chicken anemia virus coat proteins to Lactobacillus acidophilus (19), and attachment of a tumor necrosis factor alpha (TNF-␣)-specific Affibody to Lactococcus lactis (11).…”

Manipulation of Intestinal Flora as a Way to Treat Crohn's Disease: The Role of Probiotics, Prebiotics and Antibiotics

“…In another study, different binding TNF -binding molecule was applied. TNFbinding affibody was expressed and immobilized on the surface of engineered L. lactis (Ravnikar et al, 2010). As noted earlier, besides IL-10, transforming growth factor beta (TGF ) has often been described as an anti-inflammatory cytokine in CD.…”