Development of a two-step chromatography procedure that allows the purification of a high-purity anti-histone H1 monoclonal immunoglobulin M antibody with immunosuppressant activity.
In organ transplantation, the development of a novel immunosuppressant free of the need for permanent administration and any serious side effects has eagerly been awaited. We have previously reported that an anti-histone H1 polyclonal antibody has immunosuppressant activity. Here we prepared an anti-histone H1 monoclonal antibody as an analytical tool to elucidate its mechanism of immunosuppression. The isotype of this monoclonal antibody was immunoglobulin M. A monoclonal antibody prepared for administration to organ transplantation model animals should not contain any allogenic proteins and should have high purity. Therefore, we conducted a two-step chromatography procedure, consisting of strong anion-exchange chromatography and gel filtration chromatography, to purify an anti-histone H1 monoclonal immunoglobulin M antibody from the serum-free culture supernatant of hybridomas. Consequently, we successfully purified the monoclonal antibody at 96%, a purification rate at which its administration to organ transplantation model animals is possible.
Shimada
Yayoi
Y
Kazusa Institute for Drug Discovery, Josai International University, 2-1-6 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan.
Goto
Takeshi
T
Kawamoto
Seiji
S
Kiso
Takashi
T
Katayama
Akiko
A
Yamanaka
Yasushi
Y
Aki
Tsunehiro
T
Chiang
Kuei-Chen
KC
Nakano
Toshiaki
T
Goto
Shigeru
S
Chen
Chao-Long
CL
Ohmori
Naoya
N
Ono
Kazuhisa
K
Sato
Shuji
S
D000818:Animals; D000837:Anion Exchange Resins / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D018929:Cell Culture Techniques; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D016895:Culture Media, Serum-Free / Q000737:chemistry; D006657:Histones / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D006825:Hybridomas / Q000737:chemistry; D007075:Immunoglobulin M / Q000302:isolation & purification*; D007166:Immunosuppressive Agents / Q000302:isolation & purification* / Q000627:therapeutic use; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats
D000837:Anion Exchange Resins; D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free; D006657:Histones; D007075:Immunoglobulin M; D007166:Immunosuppressive Agents