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254 Galaev and Mattiasson
ensure elution of bound dog IgG and killing the residual cells which were not
washed or eluted from the column.
16. As an initial step in monitoring the binding and recovery of the cells on the
column, the absorbance measured at 470 nm (turbidity) of the fractions obtained
from the column, could be determined. The viability of the initial cell load,
the cells collected in the breakthrough fractions and cells in the eluted fractions
were checked using the Trypan blue dye exclusion method (13). The dead cells
stained dark blue and could be differentiated from the live cells, which remained
unstained. Alternatively, the cells could be labeled with fluorescent conjugated
antibodies followed by sorting and counting in a flow cytometer like FACScan
(Becton-Dickinson).
References
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(2003) Direct chromatographic capture of enzyme from crude homogenate using
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