Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria). III: gel antibodies against cells (bacteria).
Artificial antibodies in the form of gel granules were synthesized from the monomers acrylamide and N,N'-methylenebisacrylamide by the imprinting method in the presence of Echerichia coli bacteria as template. The electrophoretic migration velocities of the gel antibodies (i) saturated with the antigen (Escherichia coli MRE-600), (ii) freed of the antigen, and (iii) resaturated with bacteria, were determinated by electrophoresis in a rotating narrow-bore tube of 245 mm length and the 2.5 and 9.6 mm inner and outer diameters, respectively. Removal of bacteria from the gel antibodies was made by treatment with enzymes, followed by washing with SDS and buffer. Gel granules becoming charged by adsorption of bacteria move in an electrical field. We obtained a significant selectivity of gel antibodies for E. coli MRE-600, since the granules did not interact with Lactococcus lactis; and when E. coli BL21 bacteria were added to the gels selective for E. coli MRE-600, a significant difference in the migration rate of the complexes formed with the two strains was observed indicating the ability of differentiation between the two strains. The gel antibodies can be used repeatedly. The new imprinting method for the synthesis of artificial gel antibodies against bioparticles described herein, and the classical electrophoretic analysis technique employed, thus represent - when combined - a new approach to distinguish between different types and strains of bacteria. The application area can certainly be extended to cover other classes of cells.
Bacskay
Ivett
I
Department of Analytical Chemistry, Faculty of Science, University of Pécs, Pécs, Hungary.
Takátsy
Anikó
A
Végvári
Akos
A
Elfwing
Anders
A
Ballagi-Pordány
András
A
Kilár
Ferenc
F
Hjertén
Stellan
S
D020106:Acrylamide / Q000737:chemistry*; D000178:Acrylamides / Q000737:chemistry*; D000907:Antibodies, Bacterial / Q000737:chemistry* / Q000302:isolation & purification*; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology; D002021:Buffers; D004586:Electrophoresis / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry / Q000276:immunology; D005782:Gels / Q000737:chemistry; D015394:Molecular Structure; D011506:Proteins / Q000737:chemistry; D014780:Viruses / Q000737:chemistry
D000178:Acrylamides; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D002021:Buffers; D005782:Gels; D011506:Proteins; D020106:Acrylamide; C021221:N,N'-methylenebisacrylamide