Construction and Analysis of Recombinant Mouse Anti-Botulinum Antibodies

Anti-botulinum neurotoxin antibodies were produced by cloning heavy and light chain genes from an immunized mouse. Selection by phage display against immobilized botulinum toxin subtype B allowed for the isolation of multiple high affinity Fab producing cell lines. These clones can be directed to ex...

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Bibliographic Details
Main Authors: Emanuel, Peter A, Menking, Darrel E, Wilson, Jennifer, Valdes, James J, Dang, Jessica
Format: Report
Language:English
Subjects:
ANTIBODIES
BACTERIAL TOXINS
BACTERIOPHAGES
CELLS(BIOLOGY)
CLONES
CLOSTRIDIUM BOTULINUM
CULTURES(BIOLOGY)
DISPLAY SYSTEMS
ENGINEERING
HYBRIDOMAS
ISOLATION
LOW COSTS
Medicine and Medical Research
MICE
Microbiology
NUCLEOTIDES
PE62622A
PURIFICATION
SELECTION
SEQUENCES
SIMPLIFICATION
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Summary:Anti-botulinum neurotoxin antibodies were produced by cloning heavy and light chain genes from an immunized mouse. Selection by phage display against immobilized botulinum toxin subtype B allowed for the isolation of multiple high affinity Fab producing cell lines. These clones can be directed to express large quantities of Fab antibody at a significantly lower cost than hybridoma cell culture. Purification can be simplified by engineering an affinity tag directly into the nucleotide sequence of each antibody clone. Methods of construction and analysis are presented along with affinity constant determination and nucleotide sequencing data. Perpared in cooperation with GEO-Centers, Inc., Rockville, MD.