A model phage display subtraction method with potential for analysis of differential gene expression.
In order to establish a subtractive procedure that makes it possible to enrich selectively phage displayed antibodies directed against proteins constituting a difference between two populations of cells, a competitive selection strategy utilising two solid phases was developed and tested. Antibodies recognising a defined difference between two otherwise identical protein mixtures were isolated and their specificity confirmed. To test further the efficacy of selection inhibition during the competitive selections, selections towards a total cell extract were performed with and without competition from the same extract. An analysis of the resulting phage antibodies confirmed the subtractive nature of the system described.
Stausbøl-Grøn
B
B
Institute of Molecular and Structural Biology, Biostructural Chemistry, University of Aarhus, Denmark. groen@biobase.dk
Wind
T
T
Kjaer
S
S
Kahns
L
L
Hansen
N J
NJ
Kristensen
P
P
Clark
B F
BF
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis*; D001435:Bacteriophages; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D006801:Humans; D007135:Immunoglobulin Variable Region; D007256:Information Systems; D008545:Melanoma; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000941:Antigens; D007135:Immunoglobulin Variable Region; D011506:Proteins