Cell-free immunology: construction and in vitro expression of a PCR-based library encoding a single-chain antibody repertoire.
A novel cloning-independent strategy has been developed to generate a combinatorial library of PCR fragments encoding a murine single-chain antibody repertoire and express it directly in a cell-free system. The new approach provides an effective alternative to the techniques involving in vivo procedures of preparation and handling large libraries of antibodies. The possible use of the described strategy in the ribosome display is discussed.
Makeyev
E V
EV
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region.
Kolb
V A
VA
Spirin
A S
AS
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D015723:Gene Library; D051379:Mice; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012156:Reticulocytes / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
D000906:Antibodies; D019151:Peptide Library; D010455:Peptides; D012333:RNA, Messenger