Application of green fluorescent protein to affinity electrophoresis; affinity of IgG-binding domain C from streptococcal protein G to mouse IgG1.
Affinity electrophoresis (AEP) using green fluorescent protein (GFP) was studied. We constructed a fusion protein that linked S147PGFP and IgG binding domain C from streptococcal protein G (GFP-SpGC). The affinity of GFP-SpGC for mouse IgG1 was measured. The AEP using GFP does not require a staining step after electrophoresis, and was successful with a non-purified sample. Therefore, this method is simple and useful for measuring many samples such as those used in mutational studies.
Kazama
Hitoshi
H
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan. kazama@hoku-iryo-u.ac.jp
Yamada
Keiko
K
Aoki
Takashi
T
Watabe
Hiroyuki
H
D000818:Animals; D000915:Antibody Affinity; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D004586:Electrophoresis; D049452:Green Fluorescent Proteins; D007074:Immunoglobulin G / Q000145:classification / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins