A trinitrophenyl(TNP)-poly-L-lysine-horseradish peroxidase conjugate for the detection of anti-TNP antibodies in vivo.
A new conjugate for the detection of anti-trinitrophenyl(TNP) antibodies was developed to study the localization pattern of specific antibody containing cells and extracellular antibody in vivo. By means of a bridging molecule, poly-L-lysine, nine TNP groups and six horseradish peroxidase (HRP) groups were joined in one conjugate. Thus a higher specificity (more hapten) was united with a higher staining intensity (more enzyme) in the same conjugate. This conjugate made possible the simultaneous detection of anti-TNP antibody containing cells and establishment of their class (immunoglobulin M (IgM) and IgG). It was also used for the demonstration of anti-TNP antibodies in tissues where a TNP-alkaline phosphate (AP) conjugate could not be used due to high AP (endogenous) background staining. Thus we demonstrated anti-TNP antibody containing cells in gut associated lymphoid tissue and anti-TNP-(TNP-ovalbumin) immune complexes in the glomeruli of the kidney. We suggest that poly-L-lysine is a suitable bridging molecule for the preparation of hapten-HRP conjugates.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000918:Antibody Specificity; D005260:Female; D006735:Horseradish Peroxidase / Q000276:immunology*; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007668:Kidney / Q000032:analysis; D008198:Lymph Nodes / Q000032:analysis; D051379:Mice; D009578:Nitrobenzenes / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D010544:Peroxidases / Q000276:immunology*; D011107:Polylysine / Q000031:analogs & derivatives / Q000276:immunology*; D013154:Spleen / Q000032:analysis; D014301:Trinitrobenzenes / Q000276:immunology*
D000906:Antibodies; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D009578:Nitrobenzenes; D010455:Peptides; D014301:Trinitrobenzenes; C046207:trinitrophenyl-polylysine-horseradish peroxidase; D011107:Polylysine; D006735:Horseradish Peroxidase; D010544:Peroxidases