EMD Millipore eBook - 12

Analyzing Cell Death * Defenders Against Death

Snapshots of the selection process after each round could be obtained
by deep sequencing the ever-enriching populations. This detailed analysis
of the enrichment process allowed an interesting look at a "test tube"
selection process that pitted two replicating systems against each other.
Thus, initially a minority of cells containing protective antibodies must
compete against a majority of unprotected cells that continue to produce
large amounts of virus. n
Anton Simeonov, Ph.D., works at the NIH.

Scheme of the phenotype selection-based information enrichment. The
selection starts with a naive intrabody lentiviral library. HeLa cells were
sequentially infected with the antibody library followed by infection with
human rhinovirus. Cells expressing functional antibodies were protected
from death while other cells were killed. The integrated antibody fragments
(the information) from the surviving cells were recovered and converted into
a secondary lentiviral library that was the starting point for the next round
of selection. The selection was carried out for five rounds. The number of
surviving cells (green circles) relative to killed cells (red circle) increased in
each round. The percentage of functional antibody sequences (red and
black patches) also enriched after each round of selection.
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