상세 정보

underline
Metadata Downloads : dc(xml) or Excel
Cited 0 time in scopus ci

Title 

Bacterial expression and in vitro refolding of a single-chain fv antibody specific for human plasma apolipoprotein B-100

Authors 

Myung Hoon LeeTae In ParkYong Bok ParkJu Won Kwak

Publisher 

Elsevier

Issue Date 

2002

Citation 

Protein Expression & Purification, vol. 25, no. 1, pp. 166-173

Keywords 

apolipoprotein B-100bacterial expressionin vitro refoldingmAbB23single-chain Fvapolipoproteins Bbinding, competitivedose-response relationship, drugelectrophoresis, polyacrylamide gelenzyme-linked immunosorbent assay

Abstract 

From the cloned heavy and light chains of a murine monoclonal antibody (mAbB23) which is specific for human apolipoprotein (apo) B-100 of plasma low-density lipoproteins, a vector was designed for expression of a single-chain antibody (scFv) of mAbB23 in Escherichia coli. The expression vector was constructed so that the scFv gene (VL-linker-VH) was expressed under the control of the T7 promoter. The inclusion body of scFv was isolated from E. coli lysate and solubilized in 6 M guanidine-hydrochloride without reducing agents, followed by refolding through slow dilution into refolding buffer. After complete removal of the remaining denaturant by dialysis, the soluble scFv was purified through an apo B-100-coupled affinity column, and an active fraction, which had an antigen-binding activity comparable with that of native Fab, was easily obtained. The expression and in vitro refolding of scFv resulted in production of an active molecule in a yield of 15-20 mg per 1-liter flask cultivation.

ISSN 

1046-5928

Link 

http://dx.doi.org/10.1006/prep.2002.1623

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


There are no files associated with this item.
qrcode

FusionCharts.
DSpace Software Coptright(c) 2010 MIT and Hewleft-Packard  /  KRIBB-REPOSITORY ( Email:jakim@kribb.re.kr)