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Expression of functional antibody fragments

  • US 7,018,809 B1
  • Filed: 09/18/1992
  • Issued: 03/28/2006
  • Est. Priority Date: 09/19/1991
  • Status: Active Grant
First Claim
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1. A method for the production of a Fab′

  • antibody fragment coupled to a heterologous molecule, comprising the steps of;

    (a) expressing nucleic acid encoding a Fab′

    antibody fragment in a prokaryotic host cell culture comprising the nucleic acid such that the Fab′

    is secreted to a periplasmic space of a prokaryotic host cell;

    (b) recovering the Fab′

    from the host cell culture, wherein the recovered Fab′

    comprises a CH1 fused to the amino acid sequence of CysXX, wherein X is Ala, Arg, Asp, or Pro, and wherein the cysteine of Cys XX is present as a free thiol, and the amount of the Fab′

    recovered from the host cell culture exceeds the amount of F(ab′

    )2 recovered therefrom; and

    (c) coupling the cysteine of CysXX that is present as a free thiol of the recovered Fab′

    to a heterologous molecule.

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