W. Den et al., A bidirectional phage display vector for the selection and mass transfer of polyclonal antibody libraries, J IMMUNOL M, 222(1-2), 1999, pp. 45-57
An approach to the creation of antigen-specific polyclonal libraries of int
act antibodies is presented. A polyclonal library of Fab antibody fragments
would be expressed using a phage display vector, and selected for reactivi
ty with an antigen or group of antigens, For conversion into a sublibrary o
f intact polyclonal antibodies, the selected heavy (H) and light (L) chain
variable (V) region gene combinations would be transferred in mass, as link
ed pairs, to a eukaryotic expression vector which provides immunoglobulin (
Ig) constant (C) region genes. To enable this selection and transfer, a bid
irectional phage display vector was generated, in which the V region gene p
airs are linked head to head in opposite transcriptional orientations. The
functionality of this vector was demonstrated by the selection, transfer an
d expression of linked V region gene pairs derived from an A/J mouse that h
ad been immunized with p-azophenylarsonate (Ars)-coupled keyhole limpet hem
ocyanin (KLH). As expected, the expressed IgG2b anti-Ars antibodies with se
lected V region gene pairs were shown to have V region sequences and Ars-bi
nding characteristics similar to those of anti-Ars hybridoma antibodies. Th
e technology presented here has potential for many diagnostic and therapeut
ic applications. These include the generation of polyclonal antibody librar
ies against multiple epitopes on infectious agents or cancer cells, and of
polyclonal libraries encoding chimeric molecules composed of antibody V reg
ions and T cell receptor C regions. (C) 1999 Elsevier Science B.V. All righ
ts reserved.