Analytical control procedures of immunoreactivity for IgG and Fab fragments specific to haptens.
This study investigates immunoreactivity control procedures, i.e., specificity, affinity constant (Ka), and specific active binding sites (SABS), for polyclonal anticolchicine, monoclonal antidigitoxin IgG and Fab fragments, and antidigoxin Fab fragments (Digidot). Preliminary control procedures for IgG and Fab fragment purity indicated that all reagents were immunologically pure. All IgG and Fab fragments exhibited similar cross-reactivity and Ka. No decrease in percentage of Fab fragment SABS was observed after papain cleavage of anticolchicine and antidigitoxin IgG. Nevertheless, only 4.3 +/- 1.2% of nonimmunopurified anticolchicine polyclonal Fab fragments and 76.2 +/- 2.3 to 88.7 +/- 2.5% of different batches of immunopurified anti-digoxin Fab (Digidot) were active, the latter percentage being in the range of the 85% specified by the manufacturer. Only 58 +/- 3% of digitoxin-specific monoclonal IgG was active and 67 +/- 7% of its Fab fragments. Results show the importance of determining the ratio of SABS to presumed total specific binding sites for pharmaceutical monoclonal and polyclonal antibody preparations against haptens.
Cano
N J
NJ
INSERM U26, Hôpital F. Widal, Paris, France.
Sabouraud
A E
AE
Urtizberea
M
M
Carcagne
J
J
Grandgeorge
M
M
Scherrmann
J M
JM
D000918:Antibody Specificity / Q000276:immunology*; D001666:Binding Sites, Antibody / Q000276:immunology*; D003078:Colchicine / Q000276:immunology; D004074:Digitoxin / Q000276:immunology; D004077:Digoxin / Q000276:immunology; D006241:Haptens / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000032:analysis* / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000032:analysis* / Q000276:immunology
D006241:Haptens; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D004077:Digoxin; D004074:Digitoxin; D003078:Colchicine