mAbs are prepared by:
Monoclonal Antibody Core Laboratory
Interdisciplinary Center for Biotechnology Research
University of Florida
2033 Mowry Road
Gainesville, FL 32610
mAbs are prepared using the
CELLine 350 bioreactor flask from Integra Biosciences
BD Cell MAb basal medium from Becton Dickinson
Super low IgG fetal
bovine serum from HyClone
mAb lots are prepared as follows:
Each lot of mAb is prepared from one bioreactor flask.
Each flask is started with a vial of hybridoma cells stored at
Typically, 10-12 harvests of 8-10 mL each are obtained from one
flask over 10-12 weeks.
Harvests are sterile filtered, adjusted to 0.02% sodium azide,
and stored at 4°C.
Octet quantitation of IgG yield is done on every other harvest.
IgG concentrations are
typically in the range of 0.75 to 1.5 mg/mL.
Harvests are pooled, sterile filtered, and stored at 4°C
(pool = lot).
mAbs are tested as follows:
mAbs are tested for recognition of antigen of the correct
molecular weight by western blotting.
Western blot strips are prepared from yeast proteins separated
Blots are probed with mAbs diluted 1/1000 in PBS + 0.1% Tween
Results are visualized by chemiluminescence.
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