Selective colony blotting to expand bacterial surface receptors: applications to receptors for rat immunoglobulins.
Many bacterial surface receptors demonstrate a heterogeneous expression pattern among individual colonies. Methods have been developed to select bacteria expressing high levels of a stable surface receptor. This process is illustrated using a Streptococcus zooepidemicus isolate demonstrating a high level of Fc receptors for rat immunoglobulins. This strain was selected and expanded to obtain a bacterial isolate demonstrating approximately 100 fold greater reactivity with rat immunoglobulins than protein A positive Staphylococcus aureus or 30-40 fold higher reactivity for rat IgG than type III Fc receptor positive streptococcal group G strains. The optimal pH for rat IgG binding and the reactivity with rat IgG subclasses and certain rat monoclonal antibodies is described. The potential application and limitations of the selected rat Fc receptor positive bacterial strain to immunoassays based on the specificity of rat monoclonal and polyclonal antibodies is discussed.
Reis
K J
KJ
University of Florida.
Siden
E J
EJ
Boyle
M D
MD
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D000943:Antigens, Differentiation / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000378:metabolism*; D051381:Rats; D011961:Receptors, Fc / Q000096:biosynthesis*; D017452:Receptors, IgG; D013211:Staphylococcus aureus / Q000276:immunology*; D013291:Streptococcus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D007074:Immunoglobulin G; D011961:Receptors, Fc; D017452:Receptors, IgG