Antibody Engineering Conference, San Diego - MorphoSys
Antibody Engineering Conference, San Diego - MorphoSys
Antibody Engineering Conference, San Diego - MorphoSys
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Screening for Fab Phage Display Levels<br />
Step 1: Gene synthesis of 20 HCs and 20 LCs (including gene and codon optimization)<br />
Step 2: Pool sub-cloning into CysDisplay and Fab expression vectors<br />
Step 3: Set-up of predictive screening assays for Fab display levels on phage, Fab expression<br />
levels and Fab thermostability from crude bacterial expression samples<br />
Step 4: ELISA-based screening of HC/LC pairs; sequence analysis for identification of ≥ 75% of<br />
HC/LC pairs (> 50% at least double determination)<br />
Example: Determination of Fab phage display levels in ELISA format<br />
cultivation of E. coli<br />
carrying phagemid;<br />
helper phage infection<br />
and antibody phage<br />
production in 96-well<br />
format<br />
phage capture<br />
via anti-g8p<br />
phage capture<br />
via anti-Fab<br />
anti-g8p<br />
detection<br />
calculation of relative<br />
Fab display rates<br />
(g8p vs. Fab-specific<br />
signal) using<br />
reference phage<br />
prep<br />
Page 10 2011 <strong>Antibody</strong> <strong>Engineering</strong><br />
© <strong>MorphoSys</strong> AG
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