Improved method to raise polyclonal antibody using enhanced green fluorescent protein transgenic mice.
Recombinant fusion protein is widely used as an antigen to raise antibodies against the epitope of a target protein. However, the concomitant anticarrier antibody in resulting antiserum reduces the production of the desired antibody and brings about unwanted non-specific immune reactions. It is proposed that the carrier protein transgenic animal could be used to solve this problem. To validate this hypothesis, enhanced green fluorescent protein (EGFP) transgenic mice were produced. By immunizing the mice with fusion protein His6HAtag-EGFP, we showed that the antiserum from the transgenic mice had higher titer antibody against His6HA tag and lower titer antibody against EGFP compared with that from wild-type mice. Therefore, this report describes an improved method to raise high titer antipeptide polyclonal antibody using EGFP transgenic mice that could have application potential in antibody preparation.
Ren
Jianke
J
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Wang
Long
L
Liu
Guoxiang
G
Zhang
Wen
W
Sheng
Zhejin
Z
Wang
Zhugang
Z
Fei
Jian
J
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000276:immunology*; D006639:Histidine / Q000235:genetics / Q000276:immunology; D007114:Immunization; D007158:Immunologic Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
D000941:Antigens; D017931:DNA Primers; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006639:Histidine