Identification of dengue-specific human antibody fragments using phage display.
High-affinity antibodies are valuable tools for dengue research. A method for the selection of dengue-specific, human antibody fragments using naïve repertoires displayed on M13 filamentous bacteriophage is described. Naïve repertoires are unbiased, thus enabling the identification of antibodies to dengue structural and nonstructural proteins from the same library. Dengue-specific clones are enriched by binding to an immobilized dengue antigen, followed by washing, elution, and amplification of phage for subsequent rounds of selection. Dengue virus has four antigenically related serotypes, and the serotype of the antigen can be kept constant or alternated during the selection process depending on whether serotype-specific or cross-reactive antibodies are required. After the selection process, clones are screened, and specific clones are identified by phage ELISA and Western blot.
Emerging Infectious Diseases Program, Duke-NUS Graduate Medical School, Singapore, Singapore.
Moon Y F
MY
Tay
Chin Chin
CC
Lee
Subhash G
SG
Vasudevan
Nicole J
NJ
Moreland
D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D019921:Biotinylation; D061505:Cell Surface Display Techniques / Q000379:methods*; D003429:Cross Reactions / Q000276:immunology; D003716:Dengue Virus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D012117:Resins, Synthetic / Q000378:metabolism; D013045:Species Specificity; D019809:Streptavidin / Q000378:metabolism
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007128:Immunoglobulin Fragments; D012117:Resins, Synthetic; D019809:Streptavidin