High throughput generation and affinity maturation of humanized antibody
First Claim
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.
3 Assignments
0 Petitions
Accused Products
Abstract
Compositions, methods, and kits are provided for efficiently generating and screening humanized antibody with high affinity against a specific antigen. The library of humanized antibody is generated by mutagenizing a chimeric antibody template that combines human antibody framework and antigen binding sites of a non-human antibody. Alternatively, the library of humanized antibody is generated by grafting essential antigen-recognition segment(s) such as CDRs of the non-human antibody into the corresponding position(s) of each member of a human antibody library. This library of humanized antibody is then screened for high affinity binding toward a specific antigen in vivo in organism such as yeast or in vitro using techniques such as ribosome display or mRNA display. The overall process can be efficiently performed in a high throughput and automated manner, thus mimicking the natural process of antibody affinity maturation.
259 Citations
15 Claims
-
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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
-
Specification