Directing antigen specificity towards botulinum neurotoxin with combinatorial phage display libraries.
The production of antibodies towards antigens with low immunogenicity is enhanced by the intrinsic efficiency of screening combinatorial libraries of immunoglobulins. The need to isolate clones with rare binding specificities has dictated a highly efficient method of screening and isolating antibody clones. The production of recombinant immunoglobulin libraries in bacteria allows for a more controlled selection of antibody specificity and can be used in circumstances where hybridoma fusions are unable to isolate rare clones with the desired epitope specificity. Botulinum neurotoxin (NT) with associated non neurotoxin proteins (non-NT) as a complex was used to immunize mice to obtain mRNA for the production of a recombinant antibody library with a repertoire of specificities. Initial screens of the combinatorial library revealed clones which recognized the non-neurotoxin proteins of the toxin complex similar to monoclonal antibodies produced by conventional hybridoma fusions. The combinatorial library was re-screened in order to isolate antibodies that specifically recognized the neurotoxin component of the toxin complex. The ability to alter the biopanning selection process affords the researcher a measure of control in the selection process not available with traditional hybridoma fusions.
Emanuel
P
P
Department of Pharmacology and Experimental Therapeutics, School of Medicine, University of Maryland, Baltimore 21201, USA.
O'Brien
T
T
Burans
J
J
DasGupta
B R
BR
Valdes
J J
JJ
Eldefrawi
M
M
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D010582:Bacteriophage lambda / Q000235:genetics / Q000276:immunology*; D001905:Botulinum Toxins / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005260:Female; D015723:Gene Library; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology
D000942:Antigens, Bacterial; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins; D001905:Botulinum Toxins