Biodistribution of filamentous phage-Fab in nude mice.
In vivo panning of peptide libraries in mice has allowed the isolation of peptides which target the vasculature of specific organs. The application of this approach to phage displaying Fab fragments (phage-Fab) could lead to the isolation of antibodies which recognize novel tumor antigens. In this study, we have evaluated the biodistribution of phage-Fab in nude mice. Balb/c nude mice were injected intravenously with 10(9) TU of phage displaying the anti-colon cancer Fab c30.6. Blood samples were collected at nine time points over a period of 72 h and three groups of four mice were sacrificed at 4 min, 24 h and 72 h. Normal tissues (liver, colon, spleen, kidneys, lungs, skeletal muscle) and faeces were collected at these time points and the number of viable phage in each sample was determined. The distribution of phage in tissues was also examined by immunohistochemical analysis of paraffin-embedded tissues. Regression analysis of plasma kinetic data showed that the half-life and the volume of distribution of phage was 3.6 h and 1 ml, respectively. Phage uptake occurred predominantly in lungs, kidneys, spleen and liver. Relatively few phage were distributed to colon and muscle, and phage were eliminated from the circulation by 72 h. Immunohistochemical analysis showed phage to be mainly within the vasculature at 4 min, whereas notable phage extravasation was observed at 24 h and 72 h. In conclusion, this study provides information on the in vivo behavior of phage-Fab which will be useful in the design of in vivo panning strategies. By choosing appropriate time points for tissue collection, it may be possible to isolate novel Fabs against both intra- and extravascular targets.
Yip
Y L
YL
School of Medicine (St. Vincent's Hospital), University of NSW, Sydney, Australia.
Hawkins
N J
NJ
Smith
G
G
Ward
R L
RL
D000818:Animals; D000912:Antibodies, Neoplasm / Q000302:isolation & purification; D001435:Bacteriophages / Q000737:chemistry* / Q000276:immunology; D003110:Colonic Neoplasms / Q000276:immunology; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000097:blood / Q000378:metabolism*; D007150:Immunohistochemistry; D007700:Kinetics; D051379:Mice; D019151:Peptide Library; D014018:Tissue Distribution
D000912:Antibodies, Neoplasm; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library