Preparation of Temperature-Sensitive Antibody Fragments

摘要

We designed a temperature-sensitive single chain antibody fragment in which the antigen-binding activity was drastically altered by a temperature-shift between 4 and 37 °C. An oligonucleotide corresponding to the temperature-sensitive helix-coil transition peptide, -(-Glu Ala Ala Ala Lys-)-, was introduced between VH and VL genes of scFv against the simian virus 40 large T antigen. The antigen-binding activity of the temperature-sensitive scFv produced by Escherichia coli cells showed a maximum decrease of 1/36 at 4 °C compared with that at 37 °C. Binding activity was controlled by the NaCl concentration, as well as by the temperature shift. By using a column that immobilized the scFv, the antigen was purified in response to elution by the temperature-shift.