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An approach for preventing recombination-deletion of the 40–50 anti-digoxin antibody VHgene from the phage display vector pComb3

Authors
Journal
Gene
0378-1119
Publisher
Elsevier
Publication Date
Volume
241
Issue
1
Identifiers
DOI: 10.1016/s0378-1119(99)00462-x
Keywords
  • Anti-Digoxin Antibodies
  • Bacterial Fab Expression
  • Phage Display
Disciplines
  • Design
  • Engineering

Abstract

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.

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