С ڶءrAAVġС
ʲôڶ
ڶ(Endotoxin)ֳ֬(Lipopolysaccharide, LPS)֬AԶǺͷԽǸϸϸڵλEU/mLڶضжԵ֬AڶصҪ
ͼ1 ڶطӽṹɫʾ,ͼƬȪԴο4
rAAVΪʲôẬڶ
ڶصͷϸܻճϸʱɵҲᱬϸrAAVƱڶȾҪȪԴϸڶغڶrAAVҪDNAǴӴ˾ɢȡDNAܺڶأͬʱrAAVƱתȾ293TϸϸҲڶصıеȾڶ
rAAVеڶ
ƾ֤ҩĹ淶ҪԼڶزⶨǼڶزĽһֺ֫ɫѪҺʵɫѪҺһϸͻʹһص㿪ԼѪҺеıϸƳɺܱڶؼøԭƷͨжڶصµЧӦڶغ
ͨԼϸҺⶨж0.06λڶEU/mLƷˮϡʹⶨԼ11uL뵽rAAVҺ3760minԹܼ⽺ķԺЧ
ڶضӰ
ڶضжԵȻڶҲܻյ߷ӦڶؿϸĤϵϵ߷Ӧ֯ڵϸ֢ӵͷϸнǿĶӰϸЧڶڲ鶯ɵ»巢ȫ֢Ӧ̶ɢѪѪݿˡЧ˥յ漰ʵоվٴӦǶҪֻȥڶ
ȥrAAVڶ
ڲڶصȥͲҺϴӵҪͲȥڶؾкʱǮߡʲҺϴӹǽ˻ӽɫɢLPSɶλҺϴõŨҺķȥrAAVڶҪڲrAAVѪҪ춼һľԴͷȥڶҪ
12ꡢ30000+IJUGһŻȫϸÿ滻ϸܴһrAAVڶͬʱѡGMPɲװԴͷϿڶصıʵΡrAAVڶغ2.5EU/mlҵձ<10EU/mlΪоṩõĻûʹֿɼ
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ֻʱǻ徲
ֻʱǻʵ
ֻʱǻȹ
ڿĵط
ֻΪṩȹ̵ĸƷʲƷ
UGר
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Ӫҵѯ
UGиrAAVװŻȫƱrAAVڶغ2.5EU/mlҵ֪ٴǰٴоӪҵɨ·άϵUGǽϵ
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[1] Qingyun Zheng, et al., Low endotoxin E. coli strain-derived plasmids reduce rAAV vector-mediated immune responses both in vitro and in vivo. Mol Ther Methods Clin Dev. 2021 Jun 24;22:293-303.doi: 10.1016/j.omtm.2021.06.009.
[2]Liudmyla Kondratova, et al., Removal of Endotoxin from rAAV Samples Using a Simple Detergent-Based Protocol. Mol Ther Methods Clin Dev. 2019 Sep 6;15:112-119.doi: 10.1016/j.omtm.2019.08.013.
[3]Jihad El Andari, et al., Production, Processing, and Characterization of Synthetic AAV Gene Therapy Vectors. Biotechnol J. 2021 Jan;16(1):e2000025.doi: 10.1002/biot.202000025.
[4] Rita F Maldonado, et al., Lipopolysaccharide modification in Gram-negative bacteria during chronic infection. FEMS Microbiol Rev. 2016 Jul;40(4):480-93.doi: 10.1093/femsre/fuw007.
