Phage display and selections on cells.
Traditional methods of phage display panning bind purified antigen to plates or other solid phases to which libraries are then applied. These methods are not directly applicable to antigens in their native environment on cell surfaces or in settings where the target antigen is unknown. We describe here a procedure of a panning strategy on cell surface receptors including a depletion step. We explain every step of the protocol: production of phage library, depletion and selection, elution, screening by ELISA, and analysis of soluble antibodies by ELISA and flow cytometry. Finally, several possible variants of the protocol are explained in Subheading 4.
INSERM U624, Marseille, France.
Klervi
K
Even-Desrumeaux
Patrick
P
Chames
D000906:Antibodies / Q000378:metabolism; D001435:Bacteriophages / Q000378:metabolism; D061505:Cell Surface Display Techniques / Q000379:methods*; D002477:Cells / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology; D005434:Flow Cytometry; D006801:Humans; D012995:Solubility; D013535:Suspensions; D017186:Titrimetry
D000906:Antibodies; D013535:Suspensions