fulltext.study @t Gmail

Distinct human prolactin (hPRL) and growth hormone (hGH) behavior under bacteriophage lambda PL promoter control: Temperature plays a major role in protein yields

Paper ID Volume ID Publish Year Pages File Format Full-Text
24976 43550 2008 9 PDF Available
Title
Distinct human prolactin (hPRL) and growth hormone (hGH) behavior under bacteriophage lambda PL promoter control: Temperature plays a major role in protein yields
Abstract

When producing recombinant protein for therapy, it is desirable not only to obtain substantial amounts of the protein, but also to make sure that potential contaminants such as inducing agents are not present in the final product. To prevent this, one can use expression systems in which the promoter (λPL) is activated by a temperature shift that denatures a repressor (e.g., cIts). In this manner, hGH was successfully expressed and secreted in Escherichia coli periplasm, with specific yields well above 1 μg ml−1A600−1, after a temperature shift from 30 to 42 °C. However, attempts to express a related hormone, human prolactin, employing the same protocol were unsuccessful, providing 0.03 μg ml−1A600−1 at the most. A process is described in which this labile protein is obtained from a cIts− strain under optimized temperature condition (37 °C). The highest periplasmic secretions of prolactin ever reported were thus obtained: 0.92 ± 0.10 μg ml−1A600−1 at an optical density of ∼3 A600 units in shake flask cultures and ∼1 μg ml−1A600−1, at an OD of 35 A600 units, via a rapid and flexible batch feed process in laboratory bioreactor. Purified hPRL was monomeric, correctly processed (Mr = 22,906), properly folded and bioactive (51.5 ± 24.1 IU mg−1).

Keywords
A600, units of absorbance at a wavelength of 600 nm; CHO, Chinese hamster ovary; cIts, thermo-sensitive repressor of the PL promoter of bacteriophage λ; CRS, Chemical Reference Standard; DsbA, a 21 kDa E. coli periplasmic protein; IPTG, isopropyl-beta-d-t
First Page Preview
Distinct human prolactin (hPRL) and growth hormone (hGH) behavior under bacteriophage lambda PL promoter control: Temperature plays a major role in protein yields
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 133, Issue 1, 1 January 2008, Pages 27–35
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us