ABSTRACT
The possibility of the registration of the interaction of the cells Azospirillum lipoferum Sp59b with the specific antibodies directly in the conducting suspensions by using an acoustic sensor was shown. The main element of the sensor is a piezoelectric resonator with a lateral electric field. The analysis is based on a comparison of the resonator's electrical impedance before and after the specific biological interaction between the cells and antibodies. By using this sensor one can detect and identify the bacterial cells directly in the buffer solution with the conductivity between 2.4 and 20⯵S/cm. The minimum detectable concentration of the bacterial cells turned out to be â¼103â¯cells/ml and for a short time (less than 10â¯min). Also the possibility of the detection of the cells in the presence of the extraneous microflora was shown. The results provide the opportunities for the development of a new class of the methods for the analysis of the microbial cells in real-time directly in the buffer solution.
Subject(s)
Antibodies, Bacterial , Azospirillum lipoferum/immunology , Biosensing Techniques/methods , Acoustics , Antibody Specificity , Antigen-Antibody Reactions , Azospirillum lipoferum/ultrastructure , Buffers , Computer Systems , Electric Impedance , Lipopolysaccharides/immunologyABSTRACT
Antigenic differences were revealed between the cell wall outer membrane lipopolysaccharides and the capsular high molecular weight bioglycans for a typical strain of the nitrogen-fixing rhizobacterium Azospirillum lipoferum Sp59b using antibodies prepared against the homologous lipopolysaccharide and lipopolysaccharide-protein complex. From the capsular lipopolysaccharide-protein and polysaccharide-lipid complexes of A. lipoferum Sp59b, polysaccharides were isolated and their structure was for the first time established in Azospirillum by monosaccharide analysis which included determination of the absolute configurations, methylation, O-deacetylation, and one- and two-dimensional NMR spectroscopy. The polysaccharides of the capsular complexes were shown to have identical structure of the branched tetrasaccharide repeating unit, which differs from the structure of the O-specific polysaccharide within the outer membrane lipopolysaccharide of this strain.