Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes.
In chickens, single functional immunoglobulin variable and joining gene segments at each of the heavy and light chain loci undergo V(D)J rearrangement. Diversity is subsequently introduced by conversions templated by upstream pseudo V region genes in such a way that practically all V regions in mature B cells have identical ends. This greatly simplifies the representative amplification of V region genes. Furthermore, the entire naive repertoire of the adult chicken is produced in the bursa of Fabricius of the young bird. These special properties of the generation of immunoglobulin diversity in chickens have been exploited in the development of procedures to produce large libraries of diverse antibody combining sites derived from chicken Ig genes and expressed on filamentous bacteriophage. The utility of this library was assessed by selection of specifically binding phage using three solid phase-bound protein antigens, hen egg white lysozyme, bovine thyroglobulin and bovine serum albumin. The sequences of the V region genes thus isolated demonstrated that selection was specific and that the library contained useful diversity of binding sites. This library provides access to a repertoire whose diversity is based on a mechanism different from that underlying previously available libraries. The demonstrated feasibility of generating chicken phage antibodies may lead to the production of monoclonal reagents from immunised chickens, and the derivation of reagents for studying immunoglobulin mediated selection in avian B cell development.
Davies
E L
EL
Institute for Animal Health, Compton, Newbury, Berkshire, UK.
Smith
J S
JS
Birkett
C R
CR
Manser
J M
JM
Anderson-Dear
D V
DV
Young
J R
JR
D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D000916:Antibody Diversity; D001483:Base Sequence; D002645:Chickens / Q000276:immunology*; D017931:DNA Primers; D015723:Gene Library; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D017912:Inovirus / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D013329:Structure-Activity Relationship
D017931:DNA Primers; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D012333:RNA, Messenger; C055966:immunoglobulin Fv