التفاصيل البيبلوغرافية
العنوان:
A facile method for isolation of recombinant human apolipoprotein A-I from E. coli
المؤلفون:
Ayako Kamei , Jesse J. Tran , Robert O. Ryan , Robert S. Kiss , Jennifer A. Beckstead , Gang Ren , Paul M.M. Weers , Jennifer Shearer , Jianfang Liu , Nikita Ikon , ShiBo Feng
المصدر:
Ikon, N; Shearer, J; Liu, J; Tran, JJ; Feng, SB; Kamei, A; et al.(2017). A facile method for isolation of recombinant human apolipoprotein A-I from E. coli. Protein Expression and Purification, 134, 18-24. doi: 10.1016/j.pep.2017.03.015. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/1g08b4st
بيانات النشر:
eScholarship, University of California, 2017.
سنة النشر:
2017
مصطلحات موضوعية:
0301 basic medicine , Protein Structure , Secondary , Biochemistry & Molecular Biology , Apolipoprotein B , Active , High density lipoprotein , Phospholipid , Biological Transport, Active , Circular dichroism , Protein Structure, Secondary , Article , Protein Refolding , Thermal denaturation , law.invention , Cell Line , Reversed phase HPLC , 03 medical and health sciences , chemistry.chemical_compound , law , Escherichia coli , Humans , Denaturation (biochemistry) , Cholesterol efflux , biology , Apolipoprotein A-I , Cholesterol , Macrophages , Reverse cholesterol transport , E. coli , Biological Transport , Recombinant Proteins , 030104 developmental biology , Biochemistry , chemistry , Cell culture , biology.protein , Recombinant DNA , lipids (amino acids, peptides, and proteins) , Biochemistry and Cell Biology , Other Biological Sciences , Biotechnology , Lipoprotein
الوصف:
© 2017 Elsevier Inc. Apolipoprotein (apo) A-I is the major protein component of high-density lipoprotein (HDL) and plays key roles in the Reverse Cholesterol Transport pathway. In the past decade, reconstituted HDL (rHDL) has been employed as a therapeutic agent for treatment of atherosclerosis. The ability of rHDL to promote cholesterol efflux from peripheral cells has been documented to reduce the size of atherosclerotic plaque lesions. However, development of apoA-I rHDL-based therapeutics for human use requires a cost effective process to generate an apoA-I product that meets “Good Manufacturing Practice” standards. Methods available for production and isolation of unmodified recombinant human apoA-I at scale are cumbersome, laborious and complex. To overcome this obstacle, a streamlined two-step procedure has been devised for isolation of recombinant untagged human apoA-I from E. coli that takes advantage of its ability to re-fold to a native conformation following denaturation. Heat treatment of a sonicated E. coli supernatant fraction induced precipitation of a large proportion of host cell proteins (HCP), yielding apoA-I as the major soluble protein. Reversed-phase HPLC of this material permitted recovery of apoA-I largely free of HCP and endotoxin. Purified apoA-I possessed α-helix secondary structure, formed rHDL upon incubation with phospholipid and efficiently promoted cholesterol efflux from cholesterol loaded J774 macrophages.
وصف الملف:
application/pdf
URL الوصول:
https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4b31dc34f2293c58be1a6bd1df73306c https://escholarship.org/uc/item/1g08b4st
حقوق:
OPEN
رقم الأكسشن:
edsair.doi.dedup.....4b31dc34f2293c58be1a6bd1df73306c
قاعدة البيانات:
OpenAIRE
