The antibody variable region of isolated FGFR3-specific scFv was analyzed using phagemid vectors. The variable region sequences of heavy chain (IgG1) or light chain isolated from phagemid vectors were inserted into each mammalian expression vectors, respectively. Anti-FGFR3 antibodies were produced using the Expi293 transient mammalian expression system (Gibco, A14635, Carlsbad, CA, USA) through co-transfection of the above-mentioned vectors. Following transfection, the culture supernatant was purified using the ÄKTA protein purification system (GE Healthcare Life Sciences, Uppsala, Sweden) with HiTrap Mabselect SuRe (GE Healthcare Life Sciences, 11-0034-93, Uppsala, Sweden). After purification, enrichment was performed with Amicon® Ultra Centrifugal Filter (Merck Millipore, MA, USA). The characteristics of the highly purified antibodies were analyzed using SDS-PAGE and SEC-HPLC.
Expi293 transient mammalian expression system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Expi293 transient mammalian expression system is a convenient and efficient tool for the production of recombinant proteins in mammalian cells. It utilizes a suspension-adapted, human embryonic kidney (HEK) 293 cell line that can be easily transfected and scaled up for high-yield protein expression.
Automatically generated - may contain errors
2 protocols using expi293 transient mammalian expression system
1
Purification and Characterization of Anti-FGFR3 scFv Antibodies
2
Isolation and Modification of Anti-c-Met Antibody
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The IRCR201 antibody against human and mouse c-Met
was isolated via 6 rounds of bio-panning by the conventional method
of immobilized antigen coating on immune-tubes using the synthetic
human single chain variable fragment (scFv) phage libraries from previous
reports.18 (link),19 (link) The isolated scFv (native IRCR201) was reformatted
to full-length IgG1. A CaaX motif was inserted into the C-terminus
of the light chain of the native IRCR201 antibody sequence with a
flexible glycine linker (G7) and a CaaX motif (Cys-Val-Ile-Met) DNA
sequence at the position of cysteine 214 (Kabat numbering). The antibodies
(native IRCR201, IRCR201 and CaaX-modified IRCR201, cIRCR201) were
produced in an Expi293 transient mammalian expression system (Gibco)
and purified by a HiTrap Mabselect SuRe (GE Healthcare).
was isolated via 6 rounds of bio-panning by the conventional method
of immobilized antigen coating on immune-tubes using the synthetic
human single chain variable fragment (scFv) phage libraries from previous
reports.18 (link),19 (link) The isolated scFv (native IRCR201) was reformatted
to full-length IgG1. A CaaX motif was inserted into the C-terminus
of the light chain of the native IRCR201 antibody sequence with a
flexible glycine linker (G7) and a CaaX motif (Cys-Val-Ile-Met) DNA
sequence at the position of cysteine 214 (Kabat numbering). The antibodies
(native IRCR201, IRCR201 and CaaX-modified IRCR201, cIRCR201) were
produced in an Expi293 transient mammalian expression system (Gibco)
and purified by a HiTrap Mabselect SuRe (GE Healthcare).
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