Vector for efficient selection and/or maturation of an antibody and uses thereof
View Patent ↗It is described a vector suitable for efficient selection and/or maturation of a recombinant antibody characterized in that it contains at least one element able to reduce the expression level and/or has an improved efficiency of display of said recombinant antibody.
1. An isolated or purified vector suitable for efficient selection and affinity maturation of a recombinant antibody, the recombinant antibody having a leader peptide and being encoded by a recombinant antibody coding sequence, wherein
the vector comprises at least one element able to reduce the expression level of said recombinant antibody, the at least one element being selected from
a) a suppressed stop codon inside either the leader peptide or the recombinant antibody coding sequence;
b) a low-efficient promoter driving transcription of said recombinant antibody coding sequence; or
c) an inhibitor of the promoter driving transcription of said recombinant antibody coding sequence; and
wherein
the vector comprises a further element able to provide an efficient display of said recombinant antibody, the further element comprising:
a) a sequence coding for a carboxy-terminal part of a minor coat protein pill fused to the recombinant antibody coding sequence;
b) a leader peptide of the alkaline phosphatase of E. coli comprised as the leader peptide of the recombinant antibody; and
c) a coding sequence for the minor coat protein pill fused to the recombinant antibody coding sequence with no amber codon therebetween.
2. The vector according to claim 1 wherein the recombinant antibody includes: ScFv, active fragments of Abs, or humanized sequences of Abs.
3. The vector according to claim 1 wherein the vector is a plasmid, a phagemid or a phage.
4. The vector according to claim 1 , said vector being a phagemid vector having the nucleotide sequence of SEQ ID NO: 1.
5. An in vitro host cell transformed with the vector according to claim 1 .
6. A non-human host cell transformed with the vector according to claim 1 .
7. The non-human host cell of claim 6 , wherein the non-human host cell is a bacterial cell.
8. The vector of claim 1 , wherein the vector is a DNA vector.
9. The in vitro host cell of claim 5 , wherein the host cell is a bacterial cell.