Production, purification and characterization of an anti-(carcinoembryonic antigen) recombinant single-chain Fv antibody fragment.
Specific targeting of radioactive agents to tumour cells has been the main objective of the in vivo use of monoclonal antibodies and their fragments. In particular, specific antibodies to carcinoembryonic antigen (CEA)-expressing tumours can be used either for diagnosis or therapy, if targeting could be improved. The expression of antibody fragments in bacteria allows the preparation of engineered molecules with antigen-binding properties and a better penetration into the tumour. A specific anti-CEA single-chain Fv fragment was produced in bacteria and purified. Its binding activity has been demonstrated in ELISA, immunocytochemistry, immunohistochemistry, fluorescence-activated cell sorting and the kinetic parameters determined by the plasmon surface resonance.
Pérez
L
L
Department of Biological and Technological Research, San Raffaele Scientific Institute, Milano, Italy.
Vázquez
J E
JE
Ayala
M
M
Siccardi
A G
AG
Tresoldi
E
E
de Lalla
C
C
Gavilondo
J V
JV
Sidoli
A
A
D000912:Antibodies, Neoplasm / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D001709:Biotechnology; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D007150:Immunohistochemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D000912:Antibodies, Neoplasm; D002272:Carcinoembryonic Antigen; D017931:DNA Primers; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins