Application of protein-coupled liposomes to effective affinity screening from phage library.
For effective screening by biopanning, we propose a new affinity screening method utilizing protein-coupled liposomes (proteoliposomes) as adsorbents. With multilamellar vesicles (MLVs) composed of dipalmitoylphosphatidylcholine (DPPC): dicetylphosphate (DCP) = 10: 1 (molar ratio), adsorption of nonspecific phage VCSM13 to the liposomes without any blocking was comparable to that on polystyrene tube wall coated with blocking protein. Phages displaying octapeptides specific to an anti-peptide antibody against a peptide antigen (FVNQHLCK) were screened from an octapeptide-displayed phage library by biopanning utilizing liposomes coupled with the antibody (AB-MLVs) or a conventional immunotube coated with the antibody (AB-tube). After four rounds of biopanning, all selected phages displayed homological peptides to the antigen peptide by use of AB-MLVs, while only 15% of the selected phages displayed homological peptides in the conventional biopanning. The octapeptide selected by AB-MLVs against the anti-peptide antibody showed comparable binding affinity, which were determined by the competitive ELISA and an immunoaffinity chromatography, to that of the peptide antigen. Thus, protein-coupled liposomes are useful as adsorbents for screening from combinatorial phage libraries.
Kumada
Yoichi
Y
Graduate School of Science and Technology, Kobe University, Rokkodaicho 1-1, Nada, Kobe, Hyogo 657-8501, Japan.
Nogami
Masao
M
Minami
Naoki
N
Maehara
Masumi
M
Katoh
Shigeo
S
D000345:Affinity Labels; D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000737:chemistry*; D001483:Base Sequence; D002846:Chromatography, Affinity; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D008081:Liposomes; D011506:Proteins / Q000737:chemistry*
D000345:Affinity Labels; D017931:DNA Primers; D008081:Liposomes; D011506:Proteins