Development of a high-performance affinity chromatography-based method to study the biological interaction between whole micro-organisms and target proteins.
OBJECTIVE
The bacteria-host molecular cross-talk is the matter of primary importance both in pathogenesis and in commensalism. Principally based on immunological methods, the methodologies commonly utilized for these studies are laborious and require specific antibodies. Here, we developed a new high-performance affinity chromatography (HPAC)-based approach that allows a direct measure of the interaction between whole bacterial cells and host molecules.


RESULTS
Bifidobacterium lactis BI07 cells immobilized on amino-derivatized silica beads were utilized as stationary phase in a high-performance affinity chromatography approach. The analytes plasminogen, collagen I and collagen IV were injected, and interactions were evaluated by the insertion in an HPLC system with UV detection. According to our data, Bif. lactis BI07 is capable of interacting with plasminogen, while it does not exhibit any binding activity to collagen I and IV.


CONCLUSIONS
In this study, we implemented a high-performance affinity chromatography-based method to characterize the biological interaction between whole micro-organisms and target proteins.


CONCLUSIONS
With respect to the approaches commonly utilized to study the interaction between bacteria and host proteins, this HPAC-based approach is fast and cheaper than other methods and allows a direct measure of the interaction between bacterial cells and target molecules.
Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy.
M
M
Candela
J
J
Fiori
S
S
Dipalo
P
P
Brigidi
D001644:Bifidobacterium / Q000378:metabolism; D018914:Cells, Immobilized / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D011485:Protein Binding; D011506:Proteins / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D012822:Silicon Dioxide
D011506:Proteins; D012822:Silicon Dioxide