Evaluation of antibodies fused to minor coat protein III and major coat protein VIII of bacteriophage M13.
A gene coding for an anti-(2-phenyl-5-oxazolone) single-chain Fv antibody (Ab) fragment (anti-phOx scFv) was cloned in-frame into phagemid vectors upstream from genes encoding (i) the wild-type (wt) minor coat protein (cp) III of the filamentous bacteriophage M13 of Escherichia coli, (ii) a truncated version of cpIII (amino acid (aa) positions 198-406), (iii) the wt major cpVIII, or (iv) a hybrid of interleukin-1 beta (IL-1 beta; aa 10-152) and wt cpVIII. Recombinant (re-) phage obtained by phagemid rescue were examined for the efficiency of displaying these various anti-phOx scFv::cp hybrids with commercially available anti-M13 enzyme-linked immunosorbent assays (ELISA), by immunoblotting with an anti-c-myc Ab, and by selection experiments. We found that the highest ELISA signals were obtained with the cpIII constructs and also that more immunoreactive material was detected by blotting than with Ab::cpVIII fusions. Consequently, more scFv::cpIII than scFv::cpVIII phage could be recovered in micropanning experiments with the antigen phOx as target.
Kretzschmar
T
T
Ciba-Geigy Ltd. (K-681.5.10), Basel, Switzerland.
Geiser
M
M
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D017104:Bacteriophage M13 / Q000235:genetics*; D015153:Blotting, Western; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000276:immunology*; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010081:Oxazolone / Q000031:analogs & derivatives / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology*
D000906:Antibodies; D036022:Capsid Proteins; D007128:Immunoglobulin Fragments; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C025668:coat protein, Bacteriophage M13; C055966:immunoglobulin Fv; C018361:2-phenyloxazolone; D010081:Oxazolone