Increased yield of high purity recombinant human brain natriuretic peptide by acid hydrolysis of short fusion partner in Escherichia coli

Kanumuri, Radha Madhavi; Bajji, Chitra; Tummuru, Rajesh R.; Tatireddigari, Venkat R. R. Arva; Mangamoori, Lakshmi Narasu; Panati, Kalpana; Narala, Venkata Ramireddy

doi:10.1016/j.pep.2015.03.011

Cited by 3 publications

(3 citation statements)

References 27 publications

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“…In an attempt to answer the contradictions about size of irisin, we have designed human irisin gene according to the sequence as reported earlier [1], and purified to study its biological role. In this study, we showed the expression of irisin at 28% of total proteins, which is comparatively high in E. coli expression systems [17]. In another study, irisin was produced with a GST tag in E. coli and analysed the biological activity of GST-irisin on adipogenesis [18].…”

Section: Discussionmentioning

confidence: 80%

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Panati

Narala

Narasimha

et al. 2018

Journal of Genetic Engineering and Biotechnology

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Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of energy expenditure markers PGC-1α, UCP-1, besides abundance of beige adipocytes in WAT. Though varied sizes of irisin were reported in humans and rodents it is not yet clear about the actual size of the irisin produced physiologically. Hence, we cloned and expressed human irisin (32–143 aa of FNDC5) in Escherichia coli based on the proposed cleavage site that yields 12.5 kDa peptide to study its antigenicity and other biological functions in vitro. We purified recombinant human irisin (rh-irisin) to 95% homogeneity with simple purification method with a yield of 25 mg/g wet cell pellet. rh-irisin has been detected by commercially available antibodies from different sources with similar antigenicity. Biological activity of the rh-irisin was confirmed by using 3T3-L1 pre-adipocyte differentiation by Oil red O staining. Further, rh-irisin treatment on pre-adipocytes showed increased expression of markers associated with energy expenditure. As it is involved in energy expenditure process, it could be considered as potential therapeutic option for various metabolic diseases.

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Section: Discussionmentioning

confidence: 80%

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Panati

Narala

Narasimha

et al. 2018

Journal of Genetic Engineering and Biotechnology

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“…It has been reported that, 6 patients with AMI in combination with cardiac failure accounted for 20% to 68%, most of which were patients over 60 years of age, and patients who once developed hypertension, cardiac dilatation or recurrent cardiac infarction were at high risks of developing cardiac failure. The recent researches suggested that, 7 , 8 brain natriuretic peptide (BNP) which was a natural antagonist for renin angiotensin aldosterone system (RAAS) could produce antagonistic effect on myocardial cell, endothelin in cardiac fibroblast and vascular smooth muscle cell, norepinephrine and aldosterone. Another study has found that, 9 BNP could improve renal function and ventricular remodeling and was beneficial to the treatment of cardiac failure.…”

Section: Introductionmentioning

confidence: 99%

Effectiveness and safety of recombinant human brain natriuretic peptide in the treatment of acute myocardial infarction in elderly in combination with cardiac failure

Wang

Meng

et al. 2017

Pak J Med Sci

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Objective:To investigate the effects and safety of recombinant human brain natriuretic peptide (rhBNP) in the treatment of elderly acute myocardial infarction induced cardiac failure.Methods:One hundred and forty-six patients who were diagnosed as elderly acute myocardial infarction induced cardiac failure in the hospital from July 2014 to July 2015 were selected. They were divided into a test group and a control group, 73 each. Patients in both groups were given conventional treatment such as stabilization of atherosclerotic plaques, anti-platelet and remodeling and reversion of myocardium. The curative effects and the incidence of adverse reactions of the two groups were observed.Results:The overall efficacy of the test group and the control group was 87.7% and 65.8% respectively, and the difference had statistical significance (P<0.05). The heart rate, urine volume, n-terminal pro-brain natriuretic peptide level and left ventricular ejection fraction (LVEF) of both groups significantly improved after treatment, and the improvement of the test group was superior to that of the control group (P<0.05). The serum creatinine of the test group remarkably reduced after treatment (P<0.05). The incidence of hypotension and arrhythmia of the test group was lower than that of the control group during hospitalization period (P<0.05).Conclusion:rhBNP can effectively relieve the clinical symptoms, cardiac function indexes and hemodynamic indexes of patients with elderly acute myocardial infarction induced cardiac failure, with a high safety. It can be extensively applied in the treatment of acute myocardial infarction in combination with cardiac failure.

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“…To date, nearly 30% of currently approved recombinant therapeutic proteins are produced in E. coli . Hormones , interferons , interleukins , tumor necrosis factor alpha , cholera B subunit protein , B‐type natriuretic peptide , granulocyte colony stimulating factor , and mechano‐growth factor are among some of the approved therapeutic protein‐based products that have been produced in E. coli . Low cost and simplicity of cultivating bacteria make the E. coli expression system a preferable choice for the production of therapeutic proteins both on a laboratory scale and in industry.…”

Section: Introductionmentioning

confidence: 99%

Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

Chen

Cai

Zhang

et al. 2017

Biotech and App Biochem

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The pharmacological importance of recombinant human stem cell factor (rhSCF) has increased the demand to establish effective and large-scale production and purification processes. A good source of bioactive recombinant protein with capability of being scaled-up without losing activity has always been a challenge. The objectives of the study were the rapid and efficient pilot-scale expression and purification of rhSCF. The gene encoding stem cell factor (SCF) was cloned into pBV220 and transformed into Escherichia coli. The recombinant SCF was expressed and isolated using a procedure consisting of isolation of inclusion bodies (IBs), denaturation, and refolding followed by chromatographic steps toward purification. The yield of rhSCF reached 835.6 g/20 L, and the expression levels of rhSCF were about 33.9% of the total E. coli protein content. rhSCF was purified by isolation of IBs, denaturation, and refolding, followed by SP-Sepharose chromatography, Source 30 reversed-phase chromatography, and Q-Sepharose chromatography. This procedure was developed to isolate 5.5 g of rhSCF (99.5% purity) with specific activity at 0.96 × 10 IU/mg, endotoxin levels of pyrogen at 1.0 EU/mg, and bacterial DNA at 10 ng/mg. Pilot-scale fermentations and purifications were set up for the production of rhSCF that can be upscaled for industry.

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Increased yield of high purity recombinant human brain natriuretic peptide by acid hydrolysis of short fusion partner in Escherichia coli

Cited by 3 publications

References 27 publications

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Effectiveness and safety of recombinant human brain natriuretic peptide in the treatment of acute myocardial infarction in elderly in combination with cardiac failure

Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

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Increased yield of high purity recombinant human brain natriuretic peptide by acid hydrolysis of short fusion partner in Escherichia coli

Cited by 3 publications

References 27 publications

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Effectiveness and safety of recombinant human brain natriuretic peptide in the treatment of acute myocardial infarction in elderly in combination with cardiac failure

Large‐scale purification and characterization of recombinant human stem cell factor in Escherichia coli

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Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)

Expression, purification and biological characterisation of recombinant human irisin (12.5 kDa)