Conjugation of horseradish peroxidase to staphylococcal protein A with benzoquinone, glutaraldehyde, or periodate as cross-linking reagents.
Horseradish peroxidase was conjugated to Staphylococcal protein A by three different two-step procedures using an increasing excess of peroxidase in the second step reaction. The yield of conjugated protein A was analyzed by SDS-polyacrylamide gel electrophoresis. Conjugation of peroxidase to protein A with benzoquinone or glutaraldehyde as cross-linking reagents at a 3- to 4-fold molar excess of peroxidase resulted in a high yield of coupled protein A with conjugates of low molecular size. Conjugation of peroxidase to protein A by the periodate method resulted in a high yield of coupled protein A with polymeric conjugates of large molecular size. Based on these results, conjugates produced with glutaraldehyde as cross-linking reagents were further analyzed. The capacity of the conjugates to precipitate human immunoglobulin evaluated by radial immunodiffusion was found to be reduced to about 50% of that of native protein A. Conjugates produced with glutaraldehyde as cross-linking reagent retained 70% of the enzyme activity of native peroxidase.
D000447:Aldehydes; D016227:Benzoquinones; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005976:Glutaral; D006735:Horseradish Peroxidase; D010504:Periodic Acid; D010544:Peroxidases; D011485:Protein Binding; D011809:Quinones; D013205:Staphylococcal Protein A
D000447:Aldehydes; D016227:Benzoquinones; D003432:Cross-Linking Reagents; D011809:Quinones; D013205:Staphylococcal Protein A; D010504:Periodic Acid; C004532:quinone; D006735:Horseradish Peroxidase; D010544:Peroxidases; D005976:Glutaral