title: An approach for preventing recombination-deletion of the 40-50 anti-digoxin antibody V(H) gene from the phage display vector pComb3.
abstract: Phage display has been used extensively in antibody (Ab) engineering. Sometimes, however, phage display vectors exhibit deletion of immunoglobulin (Ig) genes. As an approach to circumvent the recombination-deletion of the murine anti-digoxin Fab 40-50 cloned into the pComb3 vector, the vector was modified with short synthetic oligonucleotides by replacing a pelB leader sequence with a gene 3 (g3) leader sequence and by using a single lacZ promoter sequence. By this means, the N-terminal amino acids of the L chain and Fd remained unchanged, and a random HCDR3 library built on this newly designed vector did not exhibit the recombination-deletion.
authors.affiliation: Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Kim
authors.forename: C C
authors.initials: CC
authors.lastname: Titlow
authors.forename: M N
authors.initials: MN
authors.lastname: Margolies
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004077:Digoxin / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D017353:Gene Deletion; D005803:Genes, Immunoglobulin / Q000235:genetics*; D005822:Genetic Vectors; D006825:Hybridomas; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D051379:Mice; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D019151:Peptide Library; D011995:Recombination, Genetic
chemical_list: D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D004077:Digoxin