scholarly journals High-performance renal imaging with a radiolabeled, non-excretable chimeric fusion protein

Theranostics ◽  
2021 ◽  
Vol 11 (18) ◽  
pp. 9177-9179
Author(s):  
Dawei Jiang ◽  
Muhsin H. Younis ◽  
Xiaoli Lan ◽  
Weibo Cai
Keyword(s):  
Fusion Protein ◽  
High Performance ◽  
Renal Imaging

scholarly journals Facilitation of Expression and Purification of an Antimicrobial Peptide by Fusion with Baculoviral Polyhedrin in Escherichia coli

2005 ◽  
Vol 71 (9) ◽  
pp. 5038-5043 ◽  
Author(s):  
Quande Wei ◽  
Young Soo Kim ◽  
Jeong Hyun Seo ◽  
Woong Sik Jang ◽  
In Hee Lee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
High Performance ◽  
Affinity Purification ◽  
Antimicrobial Activities ◽  
Host Cells ◽  
Functional Study ◽  
Fusion Partner ◽  
Expression And Purification ◽  
E Coli

ABSTRACT Several fusion strategies have been developed for the expression and purification of small antimicrobial peptides (AMPs) in recombinant bacterial expression systems. However, some of these efforts have been limited by product toxicity to host cells, product proteolysis, low expression levels, poor recovery yields, and sometimes an absence of posttranslational modifications required for biological activity. For the present work, we investigated the use of the baculoviral polyhedrin (Polh) protein as a novel fusion partner for the production of a model AMP (halocidin 18-amino-acid subunit; Hal18) in Escherichia coli. The useful solubility properties of Polh as a fusion partner facilitated the expression of the Polh-Hal18 fusion protein (∼33.6 kDa) by forming insoluble inclusion bodies in E. coli which could easily be purified by inclusion body isolation and affinity purification using the fused hexahistidine tag. The recombinant Hal18 AMP (∼2 kDa) could then be cleaved with hydroxylamine from the fusion protein and easily recovered by simple dialysis and centrifugation. This was facilitated by the fact that Polh was soluble during the alkaline cleavage reaction but became insoluble during dialysis at a neutral pH. Reverse-phase high-performance liquid chromatography was used to further purify the separated recombinant Hal18, giving a final yield of 30% with >90% purity. Importantly, recombinant and synthetic Hal18 peptides showed nearly identical antimicrobial activities against E. coli and Staphylococcus aureus, which were used as representative gram-negative and gram-positive bacteria, respectively. These results demonstrate that baculoviral Polh can provide an efficient and facile platform for the production or functional study of target AMPs.

Cystathionase: high-performance liquid chromatography. Molecular cloning in λgt11. Nonradioactive immunodetection of fusion protein

Biochimie ◽  
1989 ◽  
Vol 71 (4) ◽  
pp. 599-604 ◽  
Author(s):  
Alexander M. Shuster ◽  
Oxana A. Kvashuk ◽  
Ilia Chumakov ◽  
Vladimir S. Prassolov ◽  
Alexander G. Gabibov
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Fusion Protein ◽  
Molecular Cloning ◽  
High Performance

scholarly journals Identification of a Novel Secretogranin II-Derived Peptide (SgII187–252) in Adult and Fetal Human Adrenal Glands Using Antibodies Raised against the Human Recombinant Peptide1

1998 ◽  
Vol 83 (8) ◽  
pp. 2944-2951
Author(s):  
Youssef Anouar ◽  
Christine Desmoucelles ◽  
Laurent Yon ◽  
Jerome Leprince ◽  
Lyne Breault ◽  
...  
Keyword(s):  
Amino Acid ◽  
Adrenal Gland ◽  
Fusion Protein ◽  
Adrenal Medulla ◽  
High Performance ◽  
Polyclonal Antibodies ◽  
Physiological Role ◽  
Immunohistochemical Analysis ◽  
High Performance Liquid Chromatographic ◽  
Secretogranin Ii

abstract Molecular cloning of secretogranin II (SgII) in phylogenetically distant species has recently revealed the existence of a highly conserved 66-amino acid peptide flanked by preserved pairs of basic residues. This observation suggested that this peptide, named EM66, which had not been described to date, could be an important processing product of SgII. The aim of the present study was to investigate the possible occurrence of EM66 in the human adrenal gland. The EM66 peptide was generated in Escherichia coli, which was programmed to make a fusion protein containing the human EM66 sequence. The affinity-purified fusion protein was used to raise polyclonal antibodies in rabbits. The free EM66 peptide was obtained by cleavage of the fusion protein followed by high performance liquid chromatography purification. Immunohistochemical analysis using the EM66 antibodies revealed intense labeling of adrenochromaffin cells in the adult adrenal medulla and the fetal adrenal gland. A sensitive and specific RIA was developed and applied to the detection of EM66-like immunoreactivity in extracts of adult adrenal medulla and whole fetal adrenal gland after high performance liquid chromatographic analysis. A major immunoreactive species exhibiting the same retention time as recombinant EM66 was detected in both adult and fetal adrenal extracts. Taken together, these data demonstrate that posttranslational processing of SgII actually generates EM66 in the adrenal gland. The strong conservation of the amino acid sequence of EM66 in the vertebrate phylum and the occurrence of the mature peptide in both fetal and adult chromaffin cells suggest that EM66 could play an important physiological role in the human adrenal gland.

A High Performance Energy Analyzer for Use in Electron Scanning Microscopy

1969 ◽  
Vol 27 ◽  
pp. 14-15
Author(s):  
A. V. Crewe ◽  
M. Isaacson ◽  
D. Johnson
Keyword(s):  
High Performance ◽  
Vertical Plane ◽  
Median Plane ◽  
Electron Gun ◽  
Uniform Field ◽  
Loss Mechanism ◽  
Electron Scanning Microscopy ◽  
Sector Type ◽  
Double Focusing ◽  
Electron Energy Loss

A double focusing magnetic spectrometer has been constructed for use with a field emission electron gun scanning microscope in order to study the electron energy loss mechanism in thin specimens. It is of the uniform field sector type with curved pole pieces. The shape of the pole pieces is determined by requiring that all particles be focused to a point at the image slit (point 1). The resultant shape gives perfect focusing in the median plane (Fig. 1) and first order focusing in the vertical plane (Fig. 2).

Vacuum System to Minimize the Specimen Contamination of High-Performance EM

1977 ◽  
Vol 35 ◽  
pp. 68-69
Author(s):  
N. Yoshimura ◽  
K. Shirota ◽  
T. Etoh
Keyword(s):  
Electron Microscope ◽  
High Speed ◽  
High Performance ◽  
High Vacuum ◽  
Vacuum System ◽  
Pump System ◽  
Pumping System ◽  
Diffusion Pump ◽  
Almost All ◽  
Cascade Type

One of the most important requirements for a high-performance EM, especially an analytical EM using a fine beam probe, is to prevent specimen contamination by providing a clean high vacuum in the vicinity of the specimen. However, in almost all commercial EMs, the pressure in the vicinity of the specimen under observation is usually more than ten times higher than the pressure measured at the punping line. The EM column inevitably requires the use of greased Viton O-rings for fine movement, and specimens and films need to be exchanged frequently and several attachments may also be exchanged. For these reasons, a high speed pumping system, as well as a clean vacuum system, is now required. A newly developed electron microscope, the JEM-100CX features clean high vacuum in the vicinity of the specimen, realized by the use of a CASCADE type diffusion pump system which has been essentially improved over its predeces- sorD employed on the JEM-100C.

Use of the Magnetostatic Analogue In Electromagnetic Lens Engineering

1970 ◽  
Vol 28 ◽  
pp. 360-361
Author(s):  
John W. Coleman
Keyword(s):  
Boundary Conditions ◽  
High Performance ◽  
Force Fields ◽  
Optical Design ◽  
Direct Conversion ◽  
Design Engineering ◽  
Design Data ◽  
Scalar Potentials ◽  
Geometric Elements ◽  
At Will

In the design engineering of high performance electromagnetic lenses, the direct conversion of electron optical design data into drawings for reliable hardware is oftentimes difficult, especially in terms of how to mount parts to each other, how to tolerance dimensions, and how to specify finishes. An answer to this is in the use of magnetostatic analytics, corresponding to boundary conditions for the optical design. With such models, the magnetostatic force on a test pole along the axis may be examined, and in this way one may obtain priority listings for holding dimensions, relieving stresses, etc..The development of magnetostatic models most easily proceeds from the derivation of scalar potentials of separate geometric elements. These potentials can then be conbined at will because of the superposition characteristic of conservative force fields.

Prospects for convergent beam electron diffraction with a 200kV cold field-emission transmission electron microscope

1991 ◽  
Vol 49 ◽  
pp. 466-467
Author(s):  
J W Steeds ◽  
R Vincent
Keyword(s):  
Field Emission ◽  
High Performance ◽  
Small Diameter ◽  
Convergent Beam Electron Diffraction ◽  
Zone Axis ◽  
Medium Voltage ◽  
Transmission Electron ◽  
Good Crystallinity ◽  
Special Modification ◽  
Convergent Beam

We review the analytical powers which will become more widely available as medium voltage (200-300kV) TEMs with facilities for CBED on a nanometre scale come onto the market. Of course, high performance cold field emission STEMs have now been in operation for about twenty years, but it is only in relatively few laboratories that special modification has permitted the performance of CBED experiments. Most notable amongst these pioneering projects is the work in Arizona by Cowley and Spence and, more recently, that in Cambridge by Rodenburg and McMullan.There are a large number of potential advantages of a high intensity, small diameter, focussed probe. We discuss first the advantages for probes larger than the projected unit cell of the crystal under investigation. In this situation we are able to perform CBED on local regions of good crystallinity. Zone axis patterns often contain information which is very sensitive to thickness changes as small as 5nm. In conventional CBED, with a lOnm source, it is very likely that the information will be degraded by thickness averaging within the illuminated area.

Current State of Biological High-Resolution Scanning Electron Microscopy

1988 ◽  
Vol 46 ◽  
pp. 180-181 ◽  
Author(s):  
Klaus-Ruediger Peters
Keyword(s):  
High Performance ◽  
Low Voltage ◽  
Backscattered Electrons ◽  
Lens Position ◽  
Low Voltage Operation ◽  
Signal Collection ◽  
Probe Size ◽  
Electron Microscopes ◽  
Scanning Electron Microscopes ◽  
Scanning Electron

A new generation of high performance field emission scanning electron microscopes (FSEM) is now commercially available (JEOL 890, Hitachi S 900, ISI OS 130-F) characterized by an "in lens" position of the specimen where probe diameters are reduced and signal collection improved. Additionally, low voltage operation is extended to 1 kV. Compared to the first generation of FSEM (JE0L JSM 30, Hitachi S 800), which utilized a specimen position below the final lens, specimen size had to be reduced but useful magnification could be impressively increased in both low (1-4 kV) and high (5-40 kV) voltage operation, i.e. from 50,000 to 200,000 and 250,000 to 1,000,000 x respectively.At high accelerating voltage and magnification, contrasts on biological specimens are well characterized1 and are produced by the entering probe electrons in the outmost surface layer within -vl nm depth. Backscattered electrons produce only a background signal. Under these conditions (FIG. 1) image quality is similar to conventional TEM (FIG. 2) and only limited at magnifications >1,000,000 x by probe size (0.5 nm) or non-localization effects (%0.5 nm).

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