Abstract：
BACKGROUND:The therapeutic use of α-amidated peptides (e.g. calcitonin, glucagon-like peptide) has increased dramatically, but there are major impediments to wider use of such peptides. Larger peptides are expensive to synthesize, and short plasma half-lives frequently limit the clinical circumstances in which the peptides would be useful. Both problems are potentially solved by producing peptides as fusions with the Fc region of human immunoglobulin. METHODS:Glucagon-like peptide 1 (GLP1), peptide YY (PYY) and neuromedin U (NMU) were expressed and purified from stable CHO lines; since the α-amide group is essential for full biological potency of many peptides, Fc-fusion peptides were expressed in CHO lines stably expressing the α-amidating enzyme, peptidylglycine α-amidating monooxygenase (PAM: EC 1.14.17.3). Purified fusion proteins were analyzed intact and after HRV3C rhinovirus protease cleavage, at a site in the linker separating the Fc region from the peptide, by mass spectrometry and amide-specific immunoassays. RESULTS:The Fc fusions were expressed at 1-2.5 μg/mg cell protein and secreted at 5-20% of cell content per hour, in a peptide-specific manner. CHO cells express measurable endogenous PAM activity, amidating 25% of Fc-PYY and almost 90% of Fc-GLP1. Expression of exogenous PAM increased the level of peptide amidation to 50% of Fc-PYY and 95 % of Fc-NMU. The Fc-GLP1 fusions were 10,000-fold less active than synthetic GLP1 in a cell-receptor cyclic AMP-based assay, as expected since the amino terminal of GLP1 is essential for full biological activity. The Fc-PYY fusions were 100-fold less active than PYY-NH2 but 10-fold more active than non-amidated PYY-Gly. CONCLUSIONS:This type of approach can be used for the production of stabilized α-amidated peptides aimed at clinical trials.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Carlson KR,Pomerantz SC,Li J,Vafa O,Naso M,Strohl W,Mains RE,Eipper BAdoi
10.1186/s12896-015-0173-5subject
Has Abstractpub_date
2015-06-27 00:00:00pages
61issn
1472-6750pii
10.1186/s12896-015-0173-5journal_volume
15pub_type
杂志文章abstract：BACKGROUND:Arachidonic acid (ARA, C20:4, n-6), which belongs to the omega-6 series of polyunsaturated fatty acids and has a variety of biological activities, is commercially produced in Mortierella alpina. Dissolved oxygen or oxygen utilization efficiency is a critical factor for Mortierella alpina growth and arachidon...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-017-0388-8
更新日期：2017-08-30 00:00:00
abstract：BACKGROUND:The use of small interfering RNA (siRNA) molecules in animals to achieve double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful method of sequence-specific gene knockdown. As DNA-based expression of short hairpin RNA (shRNA) for RNAi may offer some advantages over chemical and in...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-5-13
更新日期：2005-05-11 00:00:00
abstract：BACKGROUND:Urease B is an important virulence factor that is required for Helicobacter pylori to colonise the gastric mucosa. Mouse monoclonal antibodies (mAbs) that inhibit urease B enzymatic activity will be useful as vaccines for the prevention and treatment of H. pylori infection. Here, we produced murine mAbs agai...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-10-84
更新日期：2010-11-30 00:00:00
abstract：BACKGROUND:Copper is an essential chemical element for life as it is a part of prosthetic groups of enzymes including super oxide dismutase and cytochrome c oxidase; however, it is also toxic at high concentrations. Here, we present the trade-off of copper availability and growth inhibition of a common host used for co...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-016-0251-3
更新日期：2016-02-20 00:00:00
abstract：BACKGROUND:Plasminogen activator inhibitor type-2 (PAI-2, SERPINB2) is an irreversible, specific inhibitor of the urokinase plasminogen activator (uPA). Since overexpression of uPA at the surface of cancer cells is linked to malignancy, targeting of uPA by exogenous recombinant PAI-2 has been proposed as the basis of p...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-9-43
更新日期：2009-05-14 00:00:00
abstract：BACKGROUND:Analogues of vitamin D3 are extensively used in the treatment of various illnesses, such as osteoporosis, inflammatory skin diseases, and cancer. Functional testing of new vitamin D3 analogues and formulations for improved systemic and topical administration is supported by sensitive screening methods that a...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-11-33
更新日期：2011-04-07 00:00:00
abstract：BACKGROUND:In the previous study, the cellulolytic Escherichia coli ZH-4 isolated from bovine rumen was found to show extracellular cellulase activity and could degrade cellulose in the culture. The goal of this work was to identify and characterize the secreted cellulase of E. coli ZH-4. It will be helpful to re-under...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-019-0556-0
更新日期：2019-08-27 00:00:00
abstract：BACKGROUND:The ability of cytosine deaminase (CD) to convert the antifungal agent 5-fluorocytosine (5-FC) into one of the most potent and largely used anticancer compound such as 5-fluorouracil (5-FU) raised considerable interest in this enzyme to model gene or antibody - directed enzyme-prodrug therapy (GDEPT/ADEPT) a...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-8-68
更新日期：2008-09-10 00:00:00
abstract：BACKGROUND:Cell-penetrating peptides (CPPs) have been widely used as carriers to transport different molecules into living cells, whereas messenger RNAs (mRNAs) have been utilized as target molecules for the prevention and treatment of various diseases. However, the instability of CPPs and mRNAs has limited their appli...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-016-0274-9
更新日期：2016-05-28 00:00:00
abstract：BACKGROUND:Proteins in inclusion bodies (IBs) present native-like secondary structures. However, chaotropic agents at denaturing concentrations, which are widely used for IB solubilization and subsequent refolding, unfold these secondary structures. Removal of the chaotropes frequently causes reaggregation and poor rec...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-018-0486-2
更新日期：2018-12-12 00:00:00
abstract：BACKGROUND:Plants are increasingly being examined as alternative recombinant protein expression systems. Recombinant protein expression levels in plants from Tobacco mosaic virus (TMV)-based vectors are much higher than those possible from plant promoters. However the common TMV expression vectors are costly, and at ti...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-7-52
更新日期：2007-08-27 00:00:00
abstract：BACKGROUND:Mesenchymal stem cells (MSCs) are an attractive source of adult stem cells for therapeutic application in clinical study. Genetic modification of MSCs with beneficial genes makes them more effective for therapeutic use. However, it is difficult to transduce genes into MSCs by common transfection methods, esp...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-10-38
更新日期：2010-05-13 00:00:00
abstract：BACKGROUND:β-Fructofuranosidases (or invertases) catalyse the commercially-important biotransformation of sucrose into short-chain fructooligosaccharides with wide-scale application as a prebiotic in the functional foods and pharmaceutical industries. RESULTS:We identified a β-fructofuranosidase gene (CmINV) from a Ce...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-13-100
更新日期：2013-11-14 00:00:00
abstract：BACKGROUND:Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses in the pig industry worldwide. In vivo, the virus infects a subpopulation of tissue macrophages. In vitro, PRRSV only replicates in primary pig macrophages and African green monkey kidney derived cells, such as Marc-145....
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-10-48
更新日期：2010-06-29 00:00:00
abstract：: ...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-019-0504-z
更新日期：2019-02-18 00:00:00
abstract：BACKGROUND:The identification of known mutations in a cell population is important for clinical applications and basic cancer research. In this work an immobilized form of the polymerase chain reaction, referred to as polony technology, was used to detect mutations as well as gene deletions, resulting in loss of hetero...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-3-11
更新日期：2003-07-23 00:00:00
abstract：BACKGROUND:MicroRNAs are important regulators of gene expression at the post-transcriptional level and play an important role in many biological processes. Due to the important biological role it is of great interest to quantitatively determine their expression level in different biological settings. RESULTS:We descri...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-11-70
更新日期：2011-06-25 00:00:00
abstract：BACKGROUND:Hydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materia...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-1-4
更新日期：2001-01-01 00:00:00
abstract：BACKGROUND:Interleukin-17 (IL-17), the characteristic cytokine secreted by T helper 17 lymphocytes (Th17 cells), plays a pivotal role in host defense and many inflammatory or autoimmune diseases. The aim of this study was to obtain purified protein caprine IL-17A (cIL-17A) as an antigen for preparing an IL-17A-specific...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-019-0543-5
更新日期：2019-07-17 00:00:00
abstract：BACKGROUND:The proteomics literature has seen a proliferation of publications that seek to apply the rapidly improving technology of 2D gels to study various biological systems. However, there is a dearth of systematic studies that have investigated appropriate statistical approaches to analyse the data from these expe...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-5-7
更新日期：2005-02-11 00:00:00
abstract：BACKGROUND:microRNAs (miRNAs) are short non-coding RNAs that fine-tune gene expression. The aberrant expression of miRNAs is associated with many diseases and they have both therapeutic and biomarker potential. However, our understanding of their usefulness is dependent on the tools we have to study them. Previous stud...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-018-0421-6
更新日期：2018-03-16 00:00:00
abstract：BACKGROUND:The insect cell line is a critical component in the production of recombinant proteins in the baculovirus expression system and new cell lines hold the promise of increasing both quantity and quality of protein production. RESULTS:Seventy cell lines were established by single-cell cloning from a primary cul...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-10-50
更新日期：2010-07-05 00:00:00
abstract：BACKGROUND:Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processe...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-5-18
更新日期：2005-06-16 00:00:00
abstract：BACKGROUND:Interest in using plants for production of recombinant proteins such as monoclonal antibodies is growing, but proteolytic degradation, leading to a loss of functionality and complications in downstream purification, is still a serious problem. RESULTS:In this study, we investigated the dynamics of the assem...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-11-128
更新日期：2011-12-30 00:00:00
abstract：BACKGROUND:The photorespiratory nitrogen cycle in C₃ plants involves an extensive diversion of carbon and nitrogen away from the direct pathways of assimilation. The liberated ammonia is re-assimilated, but up to 25% of the carbon may be released into the atmosphere as CO₂. Because of the loss of CO₂ and high energy co...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-11-111
更新日期：2011-11-21 00:00:00
abstract：BACKGROUND:Taxol is an effective antineoplastic agent, originally extracted from the bark of Taxus brevifolia with a low yield. Many attempts have been made to produce Taxol by chemical synthesis, semi-synthesis and plant tissue cultures. However, to date, the availability of this compound is not sufficient to satisfy ...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-6-45
更新日期：2006-12-06 00:00:00
abstract：BACKGROUND:Tetanus neurotoxin (TeNT) is taken up at nerve terminals and undergoes retrograde migration. The toxic properties of TeNT reside in the toxin light chain (L), but like complete TeNT, the TeNT heavy chain (TTH) and the C-terminal domain (TTC) alone can bind and enter into neurons. Here, we explored whether at...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-018-0452-z
更新日期：2018-06-11 00:00:00
abstract：BACKGROUND:Protein glycosylation is of fundamental importance in many biological systems. The discovery of N-glycosylation in bacteria and the functional expression of the N-oligosaccharyltransferase PglB of Campylobacter jejuni in Escherichia coli enabled the production of engineered glycoproteins and the study of the...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/1472-6750-12-67
更新日期：2012-09-24 00:00:00
abstract：BACKGROUND:Cymbidium faberi is one of the oldest cultivars of oriental orchids, with an elegant flower fragrance. In order to investigate the molecular mechanism and the functions of related proteins in the methyl jasmonate (MeJA) signaling pathway, one of the main components of flower fragrance in C. faberi, yeast one...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-020-0599-2
更新日期：2020-01-16 00:00:00
abstract：BACKGROUND:Chinese hamster ovary (CHO) cells have become the host of choice for the production of recombinant proteins, due to their capacity for correct protein folding, assembly, and posttranslational modifications. The most widely used system for recombinant proteins is the gene amplification procedure that uses the...
journal_title：BMC biotechnology
pub_type： 杂志文章
doi：10.1186/s12896-015-0218-9
更新日期：2015-10-23 00:00:00
