Targeting recombinant antibodies to the surface of Escherichia coli: fusion to a peptidoglycan associated lipoprotein.
To target recombinant antibodies to the surface of Escherichia coli, we have fused single-chain variable domains to its peptidoglycan associated lipoprotein (PAL). The fusion protein was able to bind antigen and was tightly bound to the murein layer of the cell envelope. Antibody-PAL had little effect on cell growth and viability. In contrast, the expression of single chain antibody alone eventually resulted in cell lysis. Immunofluorescence studies on unfixed cells showed that functional antibodies were accessible at the surface of intact bacteria. This could provide a means of isolating single cells producing specific antibodies from libraries in E. coli by fluorescence assisted cell sorting (FACS). Pal fusions may also be of general interest for the presentation of proteins at the surface of E. coli as, for example, in the production of live vaccines.
Fuchs
P
P
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Breitling
F
F
Dübel
S
S
Seehaus
T
T
Little
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins; D001483:Base Sequence; D001709:Biotechnology; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000276:immunology*; D029968:Escherichia coli Proteins; D005822:Genetic Vectors; D006801:Humans; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000235:genetics / Q000276:immunology*; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D004274:DNA, Recombinant; D029968:Escherichia coli Proteins; C031163:ExcC protein, E coli; D008074:Lipoproteins; D010457:Peptidoglycan; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; C069988:PplA protein, Legionella pneumophila