Regeneration of immunosorbent surfaces used in clinical, industrial and environmental biosensors. Role of covalent and non-covalent interactions.
The durability and regeneration of antibodies immobilized to commercial immunosorbents were investigated by monitoring Ag-Ab dissociation. Solutions consisting of 0.01 M hydrochloric acid (HCl), 10% propionic acid, 50% ethylene glycol and 10% SDS in 6 M urea were used in the evaluation of antigen dissociation from antibody covalently immobilized to glass and polystyrene beads, microtiter plates and Immobilon filters. RAH-IgG, used as a model antibody, bound strongly to all covalent surfaces. However, on adsorption to Nunc-1 microtiter plates, 25-60% of RAH-IgG was removed by all dissociating solutions. Covalent binding to Sanger beads was weakest relative to other covalent surfaces, exhibiting 30% and 65% detachment with ethylene glycol and SDS in urea, respectively. Although all four solutions dissociated antigen from surface-bound antibody, HCl and propionic acid were more effective on most surfaces. The antibody remained functional following antigen dissociation and reassociated to nearly 100% on all surfaces except Sanger beads and Nunc-1 microtiter plates. This study was initiated to evaluate regeneration and reuse of microelisa plates and emerging biosensors as a means of reducing routine laboratory analysis costs. Data are presented to demonstrate the reusability of microtiter plates in ELISAs following antigen dissociation from covalently bound antibody.
Veterans Administration Medical Center, Boston, MA.
G C
GC
Blanchard
C G
CG
Taylor
B R
BR
Busey
M L
ML
Williamson
D000936:Antigen-Antibody Complex; D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D006735:Horseradish Peroxidase; D007074:Immunoglobulin G; D007163:Immunosorbent Techniques; D012997:Solvents; D013464:Sulfuric Acids
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D012997:Solvents; D013464:Sulfuric Acids; D006735:Horseradish Peroxidase; C033158:sulfuric acid