Maintenance of transfection rates and physical characterization of lipid/DNA complexes after freeze-drying and rehydration

被引:76
作者
Anchordoquy, TJ [1 ]
Carpenter, JF [1 ]
Kroll, DJ [1 ]
机构
[1] UNIV COLORADO,HLTH SCI CTR,CTR PHARMACEUT BIOTECHNOL,DENVER,CO 80262
基金
美国国家科学基金会;
关键词
lyophilization; freeze-drying; cationic liposomes; stability; lipid/DNA complexes; gene delivery; transfection;
D O I
10.1006/abbi.1997.0385
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
It is well established that cationic liposomes form complexes with DNA and effectively transfect cells in vivo and ex vivo. Lipid/DNA complexes have proven safe and nonimmunogenic in clinical trials; however, they are known to aggregate readily in liquid formulations. This physical instability requires clinicians to prepare lipid/DNA complexes immediately prior to injection. In order to eliminate problems associated with this temporal requirement, we investigated the feasibility of preserving complexes as a dried preparation that could be tested, stored, and rehydrated as needed. To this end, our study evaluated the ability of different stabilizers to preserve transfection rates of complexes during acute freeze-drying stress. Our data show that complexes lyophilized in 0.5 M sucrose or trehalose possessed transfection rates similar to those of fresh preparations. In addition, dried complexes that exhibited full transfection activity upon rehydration had sizes comparable to nonlyophilized controls. Our work demonstrates that lipid/DNA complexes can be stabilized as dried powders that offer significant advantages over current liquid formulations. Furthermore, the correlation of transfection rates with maintenance of complex diameter suggests that size plays a critical role in lipid-based DNA delivery. (C) 1997 Academic Press.
引用
收藏
页码:199 / 206
页数:8
相关论文
共 44 条
[1]  
ANDERSON WF, 1995, SCI AM, V273, P124
[2]  
[Anonymous], [No title captured]
[3]   Gene therapy for cancer [J].
Blaese, RM .
SCIENTIFIC AMERICAN, 1997, 276 (06) :111-115
[4]  
Bruno M. S., 1995, Pharmaceutical Research (New York), V12, pS79
[5]   LIPOSOME-MEDIATED CFTR GENE-TRANSFER TO THE NASAL EPITHELIUM OF PATIENTS WITH CYSTIC-FIBROSIS [J].
CAPLEN, NJ ;
ALTON, EWFW ;
MIDDLETON, PG ;
DORIN, JR ;
STEVENSON, BJ ;
GAO, X ;
DURHAM, SR ;
JEFFERY, PK ;
HODSON, ME ;
COUTELLE, C ;
HUANG, L ;
PORTEOUS, DJ ;
WILLIAMSON, R ;
GEDDES, DM .
NATURE MEDICINE, 1995, 1 (01) :39-46
[6]  
Carpenter J.F., 1996, Bietechnology and biopharmaceutical manufacturing, processing, and preservation, V1996, P199
[7]   SEPARATION OF FREEZING-INDUCED AND DRYING-INDUCED DENATURATION OF LYOPHILIZED PROTEINS USING STRESS-SPECIFIC STABILIZATION .1. ENZYME-ACTIVITY AND CALORIMETRIC STUDIES [J].
CARPENTER, JF ;
PRESTRELSKI, SJ ;
ARAKAWA, T .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1993, 303 (02) :456-464
[8]  
CARPENTER JF, 1994, ACS SYM SER, V567, P134
[9]   AN INFRARED SPECTROSCOPIC STUDY OF THE INTERACTIONS OF CARBOHYDRATES WITH DRIED PROTEINS [J].
CARPENTER, JF ;
CROWE, JH .
BIOCHEMISTRY, 1989, 28 (09) :3916-3922
[10]   STABILIZATION OF DRY PHOSPHOLIPID-BILAYERS AND PROTEINS BY SUGARS [J].
CROWE, JH ;
CROWE, LM ;
CARPENTER, JF ;
WISTROM, CA .
BIOCHEMICAL JOURNAL, 1987, 242 (01) :1-10