Ke. Santora et al., Generation of a polyclonal Fab phage display library to the human breast carcinoma cell line BT-20, COMB CHEM H, 3(1), 2000, pp. 51-57
Citations number
25
Categorie Soggetti
Chemistry & Analysis
Journal title
COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING
We have previously described a vector system for generating recombinant pol
yclonal antibody libraries. This system uses bidirectional phagemid and mam
malian expression vectors to facilitate mass transfer of selected variable
light and variable heavy (VL-VH) region gene pairs from the phagemid to the
mammalian vector, to express polyclonal libraries of whole IgG antibodies.
We report here the first stage of generating a polyclonal antibody library
to the human breast carcinoma cell line BT-20, using this vector system. V
L and VH region gene pairs were obtained from a mouse immunized with BT-20
cells, and cloned, in opposite transcriptional orientations, in the bidirec
tional phagemid vector, to produce an Fab phage display library. This libra
ry was selected by panning on BT-20 cells and shown to bind specifically to
BT-20 cells. Such libraries, after suitable negative selection to eliminat
e major reactivities against normal tissue, could be transferred in mass to
our bidirectional mammalian expression vector for production of libraries
of chimeric antibodies with mouse V regions and human constant (C) regions.
These polyclonal antibody libraries will mediate effector functions and ar
e expected to be useful for breast cancer therapy, as well as diagnosis.