An approach for preventing recombination-deletion of the 40-50 anti-digoxin antibody V(H) gene from the phage display vector pComb3.
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.
Kim
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SH
Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
Titlow
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CC
Margolies
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MN
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
D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D004077:Digoxin