A New Assay Using Surface Plasmon Resonance (SPR) to Determine Binding of theLactobacillus acidophilusGroup to Human Colonic Mucin

Abstract

A new binding assay to investigate the mechanism of adhesion of lactic acid bacteria to the human intestine was established by the surface plasmon resonance technique using a biosensor BIACORE1000. Cells of 26 strains of the Lactobacillus acidophilus group as analytes were eluted onto a sensor chip on which were immobilized biotinylated A-trisaccharide polymer probes having human A-type antigen [(GalNAcalpha1-3(Fucalpha1-2)Gal)-] or human colonic mucin of blood type A (HCM-A) as ligands. In the first screening, high adhesive affinity to the A-trisaccharide BP-probe was observed in L. acidophilus OLL2769, L. crispatus JCM8778, LA205 and LA206. In the second screening, which used HCM-A, only L. acidophilus OLL2769 and L. crispatus JCM8778 were selected as adhesive strains with specific binding ability to human A-antigen. The results indicated that some strains of the L. acidophilus group could recognize and bind the sugar chain of A-antigen structure on HCM.

An Adhesin-Like Protein, Lam29, fromLactobacillus mucosaeME-340 Binds to Histone H3 and Blood Group Antigens in Human Colonic Mucus

“…The interaction of analyte (ME-340 cells) and ligand (human recombinant histone H3 or BSA-A) was evaluated on a biosensor BIACORE 1000 (GE Healthcare Bio-Sciences KK, Tokyo, Japan), which uses quantitative surface plasmon resonance (SPR), as described previously. 17) Histone H3 protein was immobilized on a CM5 research grade chip (GE Healthcare Bio-Sciences KK) using amine coupling following to the manufacturer's instructions. The analyte was suspended (0.1 mg/mL) in HBS-EP buffer (HEPES buffered saline with EDTA and polysorbate 20: 0.01 M HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.005% surfactant P20; GE Healthcare Bio-Sciences KK) and injected at a flow rate of 3 mL/min at 25 C for 5 min.…”

Evaluation of bifidobacterial adhesion to acidic sugar chains of porcine colonic mucins

“…SPR-based adhesion assays were performed using a Biacore-X system (GE Healthcare) according to the method described by Uchida et al, 24) with the following modifications. Bacterial cells were suspended in HBS-EP buffer (10 mM HEPES, 0.15 M NaCl, 3.4 mM EDTA, 0.005% surfactant P20, pH 7.2) at a concentration of 1 × 10 8 colony-forming units (cfu)/mL.…”

Lactic Acid Bacteria (LAB) Bind to Human B- or H-Antigens Expressed on Intestinal Mucosa

“…Hideaki UCHIDA, 1;y Yasushi KAWAI, 1 Hideki KINOSHITA, 1 Haruki KITAZAWA, Twenty lactobacilli isolated from human feces were studied for binding to the human blood type B-antigen [Gal1-3 (Fuc1-2) Gal-] and H-antigen (Fuc1-2Gal-] expressed sugar chains in human intestinal mucosa. We found two strains, L. gasseri OLL2755 and L. gasseri OLL2877 that firmly adhere to human B-antigen, and we found L. gasseri OLL2827 bound to the H-antigen.…”