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High throughput generation and affinity maturation of humanized antibody

  • US 10,329,555 B2
  • Filed: 02/22/2016
  • Issued: 06/25/2019
  • Est. Priority Date: 08/12/2002
  • Status: Active Grant
First Claim
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1. A method for screening a library of human or humanized antibodies in yeast, comprising:

  • (i) assembling a library of antibodies in yeast cells, wherein the step of assembling the library comprises;

    (a) mutagenizing at least one of a VH and a VL to produce a plurality of VH and/or VL; and

    (b) introducing the plurality of VH and/or VL into an expression vector by homologous recombination;

    (ii) expressing the library of antibodies in the yeast cells, wherein each of the antibodies comprises a VH and a VL that form a VH/VL heterodimer in the yeast cells;

    (iii) contacting a specific target protein with the library of antibodies expressed in the yeast cells; and

    (iv) selecting those antibodies based on their binding to the target protein.

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