A family of vectors for surface display and production of antibodies.
Expression vectors for surface display and production of single-chain (Fv) antibodies (scAb) have been constructed based on the phagemid pSEX, which expresses DNA encoding a scAb fused to the gene III product of filamentous phage [Breitling et al., Gene 104 (1991) 147-153]. A smaller version of this phagemid, pSEX20, was made by removing an unnecessary cat. To produce a vector for the surface display of other proteins and peptides, the scAb of pSEX20 was substituted by a polycloning site (MCS) to give pSEX40. For the presentation of Ab on the surface of Escherichia coli, phagemid pAP10 was derived from pSEX20 by substituting gene III with a gene encoding the peptidoglycan-associated lipoprotein (PAL). Vectors for producing scAb that can be purified by antibody and metal affinity chromatography were constructed by substituting gene III in the vector pSEX20 with DNA encoding a peptide with a C-terminal epitope recognised by a monoclonal antibody (phagemid pOPE40) or with five C-terminal histidines (pOPE 90).
Dübel
S
S
Division of Diagnosis and Experimental Therapy, German Cancer Research Center, Heidelberg.
Breitling
F
F
Fuchs
P
P
Braunagel
M
M
Klewinghaus
I
I
Little
M
M
D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D017123:Bacteriophage T7 / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015183:Restriction Mapping; D013728:Terminator Regions, Genetic; D014169:Transformation, Bacterial
D000906:Antibodies; D000939:Epitopes; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins