Semisynthetic combinatorial antibody libraries: a chemical solution to the diversity problem

Proc Natl Acad Sci U S A. 1992 May 15;89(10):4457-61. doi: 10.1073/pnas.89.10.4457.

Abstract

The properties of naiveté and large diversity are considered to be essential starting features for combinatorial antibody libraries that eschew immunization by evolution in vitro. We have prepared large libraries with such properties by using random oligonucleotide synthesis, which has the potential to create approximately 10(20) complementarity-determining regions for antibody heavy chains. When combined with light chains and expressed on phage surfaces, high-affinity antibodies could be selected from 5.0 x 10(7) Escherichia coli transformants. Remarkably, antibodies selected only for binding displayed both general structural features known to be important in nature's own antibodies and specific consensus sequences thought to be critical for interaction with the hapten against which the library was selected. Semisynthetic and ultimately totally synthetic combinatorial libraries when coupled with mutation and selection procedures should replace immunization for generation of reagent, therapeutic, and catalytic antibodies.

MeSH terms

  • Amino Acid Sequence
  • Antibody Diversity / genetics*
  • Antibody Specificity
  • Base Sequence
  • Binding Sites, Antibody
  • Binding, Competitive
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / genetics
  • Gene Library
  • Genetic Variation*
  • Humans
  • Immunoglobulin Fab Fragments / genetics*
  • Immunoglobulin Heavy Chains / genetics*
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / chemical synthesis*
  • Plasmids

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin Heavy Chains
  • Oligodeoxyribonucleotides