Chimaeric phages
The invention relates to the field of generating helper phages and phage display libraries for the identification of binding molecules. The invention provide chimaeric phages having a coat comprising a protein mixture. The protein mixture comprises a fusion protein having a proteinaceous molecule fused to a functional form of a phage coat protein and a mutant form of the phage coat protein, wherein the mutant form is impaired in binding to a host cell receptor. The invention further provides new phage collections, novel helper phages and methods and means for producing chimaeric phages, infectious phages and helper phages.
1. A chimaeric phage displaying a mixture of proteins in its coat, said mixture of proteins comprising:
a fusion protein comprising a proteinaceous molecule fused to the g3 phage coat protein; and
a mutant form of said g3 phage coat protein,
wherein the D1 region, D2 region, or both the D1 and the D2 regions of the g3 protein are deleted from said mutant form of the g3 phage coat protein, wherein the mutant form comprises the D3 region, and wherein the chimaeric phage's genomic DNA sequences are based on the genomic sequences said phage of a M13, M13K07, VCSM13, or R408 phage.
2. The chimaeric phage of claim 1 , further comprising a nucleic acid encoding said fusion protein.
3. The chimaeric phage of claim 1 , wherein said proteinaceous molecule is selected from the group consisting of an antibody, a Fab fragment, a single chain Fv fragment, a variable region, a CDR region, and an immunoglobulin.
4. The chimaeric phage of claim 2 , wherein expression of said fusion protein is under the control of a regulatable promoter.
5. The chimaeric phage of claim 4 , wherein said promoter is selected from the group consisting of a AraC/BAD promoter, a psp promoter and a lac promoter.
6. A phage collection comprising the chimaeric phage of claim 1 .
7. The phage collection of claim 6 , wherein said phage collection is a phage display library.
8. A phage collection consisting essentially of chimaeric phages according to claim 1 .