Preparative electroelution of specific protein antigens from Mycoplasma pneumoniae: use in an enzyme-linked immunosorbent assay (ELISA).
A rapid, simple and preparative method is described for the recovery of the seven highest molecular weight proteins (HMWP) from Mycoplasma pneumoniae membrane. The yield of proteins obtained was approximately 90%. The method involved the separation of M. pneumoniae proteins by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), followed by electroelution of HMWP. These eluted antigens were used in an ELISA to measure IgG antibodies in sera from 9 blood donors and 9 patients with M. pneumoniae infection. The specificity of M. pneumoniae HMWP was examined by competition ELISA and immunoblotting with different mycoplasma species encountered in the respiratory tract.
Papierok
G
G
Service de Microbiologie, Institut Pasteur, Lille, France.
Defives
C
C
Daunizeau
A
A
Wattré
P
P
Derieux
J C
JC
D000907:Antibodies, Bacterial / Q000032:analysis*; D000942:Antigens, Bacterial / Q000276:immunology / Q000302:isolation & purification*; D002462:Cell Membrane / Q000276:immunology; D003168:Complement Fixation Tests; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000032:analysis; D008970:Molecular Weight; D009177:Mycoplasma pneumoniae / Q000276:immunology*; D011019:Pneumonia, Mycoplasma / Q000276:immunology*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007074:Immunoglobulin G