Rapid Immunocapture of Pseudomonas putida Cells from Lake Water by Using Bacterial Flagella.

摘要: Abstract Monoclonal antibodies to Pseudomonas putida Paw340 cells were produced. In an enzyme-linked immunosorbent assay (ELISA) against whole bacterial cells, a hybridoma cell line termed MLV1 produced monoclonal antibody that reacted with P. but showed no cross-reaction 100 medical isolates and 150 aquatic isolates. By ELISA, immunogold electron microscopy, Western blot (immunoblot) analysis, was found react purified flagella. The surfaces of magnetic polystyrene beads coated antibody. mixing antibody-coated lake water samples containing the target host, bead-cell complexes which could be recovered by attraction towards magnet formed. Prevention nonspecific attachment required incorporation detergents in isolation protocol. These affected colony-forming ability; however, remained intact for direct detection. When reisolated standard cultural methods, approximately 20% initial population recovered. Since field, bacteria separated from other organisms particulate material not attracted thereby enriched. This method may now provide useful system recovering recombinant selectively environmental samples.