Changes in the specificity of antibodies against steroid antigens by introduction of mutations into complementarity-determining regions of the V(H) domain

Yoshitaka Iba, Nobuhiro Hayashi, Jun Ichi Sawada, Koiti Titani, Yoshikazu Kurosawa

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22 Citations (Scopus)

Abstract

An attempt was made to change the specificity of antibodies (Abs) by introduction of mutations. A monoclonal Ab specific for 17α-hydroxyprogesterone (17-OHP) was used as the starting Ab. On the basis of a model that was generated by a computer-driven model-building system, we constructed a phage-display library of Abs in which 16 residues were mutated in three complementarity-determining regions of the heavy chain that appeared to form the steroid-binding pocket. We screened the library with 17-OHP and cortisol that had been conjugated with bovine serum albumin, and we isolated many clones that had retained 17-OHP-binding ability as well as clones with the newly developed ability to bind cortisol in addition to 17-OHP. We compared the amino acid sequences between 17-OHP-specific and cortisol-binding Abs, and then constructed several additional Abs. Our results indicated that a change in specificity could be achieved by changing only a single, critical amino acid residue. Models of the 17-OHP- and cortisol-binding pockets formed by the mutated Abs could explain these observations.

Original languageEnglish
Pages (from-to)361-369,370
JournalProtein Engineering
Volume11
Issue number5
DOIs
Publication statusPublished - 05-1998

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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