Focused libraries of genetic packages

• US 8,895,475 B2
• Filed: 04/16/2010
• Issued: 11/25/2014
• Est. Priority Date: 12/18/2000
• Status: Expired due to Term

• Current Assignee: Dyax Corporation (Shire plc)
• Original Assignee: Dyax Corporation (Shire plc)
• Inventors: Ladner, Robert Charles
• Primary Examiner(s): BOESEN, CHRISTIAN C
• Application Number:
  US12/762,051
• Publication Number:
  US 20100292103A1
• Time in Patent Office: 1,684 Days
• Field of Search: None
• US Class Current: 506/17
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/005 constructed by phage libraries
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C40B 40/02 Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
• CPC Class Codes: C40B 40/08 Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof

Abstract

Focused libraries of vectors or genetic packages that display, display and express, or comprise a member of a diverse family of antibody peptides, polypeptides or proteins and collectively display, display and express, or comprise at least a portion of the focused diversity of the family. The libraries have length and sequence diversities that mimic that found in native human antibodies.

Claims

1. A focused library of vectors or genetic packages, the focused library comprising vectors or genetic packages which are characterized by variegated DNAs, each of which comprises a sequence that encodes a human antibody heavy chain comprising a synthetic heavy chain CDR1, CDR2 and CDR3,

wherein the human antibody heavy chain comprises a human heavy chain variable region, wherein the heavy chain variable region consists of a sequence represented by the formula V:

;

nz;

;

D;

;

ny;

;

JHn, wherein V is a v gene, nz is a series of bases that are essentially random, D is a D segment, ny is a series of bases that are essentially random, and JHn is one of the six JH segments;

wherein the v gene is 3-23, 4-34, 3-30, 3-30.3, or 4-30.1;

wherein JH is JH4 or JH3;

wherein the CDR1 segments encoded by the variegated DNAs comprise CDR1 segments that consist of a length of 5 amino acid residues;

wherein the CDR2 segments encoded by the variegated DNAs comprise CDR2 segments that consist of a length of 17 amino acid residues;

wherein the CDR3 segments encoded by the variegated DNAs comprise an embedded human antibody heavy chain D segment, wherein the CDR3 segments consist of a length of 7-15 amino acid residues between YYCAR and WG;

wherein the CDR3 segments in the library comprise amino acid sequence variation and amino acid length variation;

wherein the variegation is focused so as to closely mimic the naturally-occurring diversity of the specific family that the library represents, both in sequence and in length.

2. The focused library of vectors or genetic packages of claim 1, further comprising vectors or genetic packages which are characterized by variegated DNA sequences that encode a light chain comprising CDR1, CDR2 and CDR3, the light chain being a kappa light chain.

3. The library of claim 2, wherein the library is a library of yeast vectors.

4. The library of claim 1, wherein the library is a library of genetic packages.

5. The library of claim 1, wherein the library is a library of yeast vectors.

6. The focused library of claim 1, wherein the v gene is 3-23 and the JH is JH4.

7. The focused library of claim 1, wherein the synthetic heavy chain CDR1 segments encoded by the variegated DNAs comprise amino acid sequence variation.

8. The focused library of claim 1, wherein the synthetic heavy chain CDR2 segments encoded by the variegated DNAs comprise amino acid sequence variation.

9. The focused library of claim 1, wherein the synthetic heavy chain CDR1 segments further comprise lengths of 7 amino acid residues.

10. The focused library of claim 1, wherein the synthetic heavy chain CDR1 segments further comprise lengths of 14 amino acid residues.

11. The focused library of claim 1, wherein the synthetic heavy chain CDR2 segments further comprise lengths of 16 amino acid residues.

12. The focused library of claim 1, wherein the synthetic heavy chain CDR2 segments further comprise lengths 19 amino acid residues.

13. The library of claim 2, wherein the library is a library of genetic packages.