Abstract:
:Midkine is a heparin-binding growth factor that promotes cell growth, survival, and migration. Externally added midkine prevents ventricular remodeling and improves long-term survival after myocardial infarction in the mouse. Preclinical testing of this protein is in progress. Externally added pleiotrophin, a member of the midkine protein family, promotes functional recovery after neural transplantation in rats. Thus, pleiotrophin is also a candidate therapeutic protein. Large amounts of these proteins were obtained by using the heterologous protein expression system of Pichia pastoris, and the recombinant P. pastoris clones were cultured in a controlled fermentor. Intracellular expression yielded about 300 mg/L recombinant human (rh)-midkine, which was extracted, renatured, and purified. From 1 L of the culture, 64 mg of rh-midkine was purified. Secretory expression induced by the midkine secretion signal resulted in about 100 mg of rhmidkine in 1 L of the culture supernatant, but over 70% of the rh-midkine had yeast-specific glycosylation. Three threonyl residues that are targets for glycosylation were substituted with alanyl residues, and nonglycosylated, active rh-midkine was obtained. In secretory expression using α-mating factor prepro-sequence, about 640 mg/L rh-midkine was obtained, but it was partially truncated. Therefore, a protease-deficient host was used, and about 360 mg/L intact rh-midkine was then obtained. The rh-midkine was recovered and purified, with 70% final yield. All purified rh-midkine, regardless of expression method, was able to promote mammalian cell proliferation. In secretory expression of rh-pleiotrophin using α- mating factor prepro-sequence, 260 mg/L rh-pleiotrophin could be secreted. The rh-pleiotrophin was recovered and efficiently purified with 72% final yield.
journal_name
Curr Pharm Biotechnoljournal_title
Current pharmaceutical biotechnologyauthors
Murasugi Adoi
10.2174/1389201014666131226114919subject
Has Abstractpub_date
2013-01-01 00:00:00pages
768-84issue
8eissn
1389-2010issn
1873-4316pii
CPB-EPUB-58284journal_volume
14pub_type
杂志文章abstract:BACKGROUND:L2-based Human Papillomavirus (HPV) prophylactic vaccines, containing epitopes from HPV minor capsid proteins, are under investigation as second-generation HPV vaccines. No such vaccine has passed clinical trials yet, mainly due to the low immunogenicity of peptide vaccines; so efforts are being continued. A...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/1389201020666191114104850
更新日期:2020-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
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journal_title:Current pharmaceutical biotechnology
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journal_title:Current pharmaceutical biotechnology
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更新日期:2018-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/1389201014666131226160025
更新日期:2013-01-01 00:00:00
abstract:: ...
journal_title:Current pharmaceutical biotechnology
pub_type: 社论
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更新日期:2019-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
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journal_title:Current pharmaceutical biotechnology
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更新日期:2020-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/1389201018666170519095339
更新日期:2017-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
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更新日期:2011-04-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920111798281045
更新日期:2011-09-01 00:00:00
abstract::After a tooth extraction both hard and soft tissues undergo dimensional changes. The use of a ridge preservation technique at the moment of the tooth extraction could help could help to prevent the volume loss, thus simplifying the subsequent prosthetic and/or implant therapy. Some studies were conducted in order to e...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/1389201018666161223152607
更新日期:2017-01-01 00:00:00
abstract::The accidental or intentional release of plutonium or americium can cause acute and long term adverse health effects if they enter the human body by ingestion, inhalation, or injection. These effects can be prevented by rapid removal of these radionuclides by chelators such as calcium or zinc diethylenetriaminepentaac...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920112802273308
更新日期:2012-08-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920111795909113
更新日期:2011-07-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
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更新日期:2012-05-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920106776597612
更新日期:2006-04-01 00:00:00
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更新日期:2014-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/1389201020666190222185209
更新日期:2019-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920111798808400
更新日期:2011-12-01 00:00:00
abstract::This mini-review describes how single-molecule sensitive fluorescence resonance energy transfer (FRET) and photoinduced electron transfer (PET) reactions can be successfully applied to monitor conformational dynamics in biopolymers. Single-pair FRET experiments are ideally suited to study conformational dynamics occur...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/1389201043376896
更新日期:2004-06-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/1389201021666201203160012
更新日期:2020-12-03 00:00:00
abstract::Nitric oxide (NO) participates in the control of the cardiovascular system where two constitutive isoforms of NO-synthase were discovered: endothelial and neuronal. Both isoforms were observed in various cells, however, endothelial NO-synthase is predominantly present in the endothelium. Injury of the endothelium dist...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920111798280992
更新日期:2011-09-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/1389201018666170127105555
更新日期:2017-01-01 00:00:00
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journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章,评审
doi:10.2174/138920106777950834
更新日期:2006-08-01 00:00:00
abstract::In order to valorize novel biotechnology innovations, there is a need to evaluate ex-ante their market potential. A case in point is biofortification, i.e. the enhancement of the micronutrient content of staple crops through conventional or genetic breeding techniques. In a recent article in Nature Biotechnology, for ...
journal_title:Current pharmaceutical biotechnology
pub_type: 杂志文章
doi:10.2174/138920112804724873
更新日期:2012-12-01 00:00:00