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Phage‐displayed Fab fragments against anti‐human interleukin‐2 receptor α. Detection of antigen‐bound phages with anti‐cpIII monoclonal antibodies
Author(s) -
RAIVIO EVA,
KRONQVIST MALIN,
BRODIN THOMAS N.,
PERSSON MATS A. A.,
LINDQVIST CHRISTER
Publication year - 1997
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1111/j.1699-0463.1997.tb00548.x
Subject(s) - monoclonal antibody , antigen , antibody , virology , phage display , monoclonal , immunology , microbiology and biotechnology , biology
The genes encoding the V H C H1 and V L C L parts of the mouse anti‐human IL‐2Rα antibody 7G7B6 were amplified by PCR and the corresponding antibody fragments displayed on the surface of filamentous phages. The expression of Fab fragments was analysed by immunoblotting using HRP‐labelled goat anti‐mouse Ig antisera. By traditional hybridoma technology, splenocytes from Balb/c mice, immunized with native phage particles, were fused with P3X63‐Ag8.653 myeloma cells in order to yield monoclonal antibodies against filamentous phage proteins. The obtained monoclonal antibody IF8 (μ/ k ) recognized the minor coat protein III as a 65–70 kDa protein band by immunoblotting, whereas the monoclonal antibody IVC8 (μ/ k ), in addition to cpIII, recognized a protein with an approximate molecular weight of 38–43 kDa. Both antibodies were employed to determine the binding specificity of the phage‐displayed anti‐human IL‐2Rα Fab fragments in an ELISA using recombinant baculovirus‐expressed human IL‐2Rα proteins as antigens.