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High Range Porcine Insulin ELISA(1x96wells)

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[#10-1223-01] High Range Porcine Insulin ELISA(1x96wells)

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(1) A novel potential primary method for quantification of enantiomers by high performance liquid chromatography-circular dichroism.[TOP]

Pubmed ID :29743524
Publication Date : //
Primary methods play an important role in metrology. They can be used for the value assignment of certified reference materials, enabling the accuracy and comparability of the measurement. A novel potential primary method for enantiomer quantitation based on high-performance liquid chromatography-circular dichroism is described using L-phenylalanine as an example. The optimal quantitation range of L-Phe was from 0.1 mg/g to 1.2 mg/g, where both the relative bias and method variance were lower than 1%. The LOD and LOQ were 4 μg/g and 30 μg/g, respectively. The proposed method was also applied to the determination of the mass fraction of pure porcine insulin in solid. The average mass fraction obtained was 0.922 g/g with a RSD of 1.5%, and the associated relative uncertainty is 3.8% (k = 2), which agreed well with that obtained from the traditional isotope dilution mass spectrometry method. The LOD and LOQ for insulin quantitation were found to be 0.12 mg/g and 0.44 mg/g, respectively. The proposed method can be entirely described and understood by equations and a complete uncertainty statement can be defined in SI units.Therefore, it may be a potential primary method useful for the quantification of chiral compounds and proteins, and a supplementary method to the traditional isotope dilution mass spectrometry approach.

Authors : Luo Yi, Wu Liqing, Yang Bin, Jin Youxun, Zheng Kangle, He Zhangjing,



(2) A validated UPLC-MS/MS method coupled with protein precipitation and ion exchange solid phase extraction for the quantitation of porcine relaxin B29 in dog plasma and its application to a pharmacokinetic study.[TOP]

Pubmed ID :28889219
Publication Date : //
Porcine relaxin is a 6 kDa peptide hormone of pregnancy with important physiological and pharmacological effects. It contains a number of analogs of which porcine relaxin B29 is one of the most important. To support the development of porcine relaxin B29 as a new drug, we established an UPLC-MS/MS method for its quantitation in dog plasma. Sample preparation by protein precipitation and ion exchange solid phase extraction was followed by UPLC on an XBridge™ BEH300 C18 column at 40 °C in a run time of only 5.5 min. Detection was performed on a Qtrap 6500 mass spectrometer using ESI in the positive ion mode with MRM of the transitions at m/z 831.7 [M+7H] → 505.4 and m/z 1162.4 [M+5H] → 226 for pRLX B29 and internal standard (recombinant human insulin), respectively. The method was linear over the concentration range 30-2000 ng/mL with no matrix effects. Intra- and inter-day precisions were < 15% with accuracies in the range 98.8-100.6%. The method was successfully applied to a pharmacokinetic study in beagle dogs after administration of a 0.15 mg/kg intravenous dose. Graphical abstract Sample preparation and detection procedure.

Authors : Su Chong, Sun Hui, Yang Hong, Yin Lei, Zhang Jiwen, Fawcett John Paul, Gu Jingkai,



(3) Expressed alleles of imprinted IGF2, DLK1 and MEG3 colocalize in 3D-preserved nuclei of porcine fetal cells.[TOP]

Pubmed ID :27716032
Publication Date : //
To explore the relationship between spatial genome organization and gene expression in the interphase nucleus, we used a genomic imprinting model, which offers parental-specific gene expression. Using 3D FISH in porcine fetal liver cells, we compared the nuclear organization of the two parental alleles (expressed or not) of insulin-like growth factor 2 (IGF2), a paternally imprinted gene located on chromosome 2. We investigated whether its nuclear positioning favors specific locus associations. We also tested whether IGF2 is implicated in long-range chromatin trans-associations as previously shown in the mouse model species for its reciprocal imprinted gene H19.

Authors : Lahbib-Mansais Yvette, Barasc Harmonie, Marti-Marimon Maria, Mompart Florence, Iannuccelli Eddie, Robelin David, Riquet Juliette, Yerle-Bouissou Martine,



(4) Cavitation-enhanced delivery of insulin in agar and porcine models of human skin.[TOP]

Pubmed ID :25716689
Publication Date : //
Ultrasound-assisted transdermal insulin delivery offers a less painful and less invasive alternative to subcutaneous insulin injections. However, ultrasound-based drug delivery, otherwise known as sonophoresis, is a highly variable phenomenon, in part dependent on cavitation. The aim of the current work is to investigate the role of cavitation in transdermal insulin delivery. Fluorescently stained, soluble Actrapid insulin was placed on the surface of human skin-mimicking materials subjected to 265 kHz, 10% duty cycle focused ultrasound. A confocally and coaxially aligned 5 MHz broadband ultrasound transducer was used to detect cavitation. Two different skin models were used. The first model, 3% agar hydrogel, was insonated with a range of pressures (0.25-1.40 MPa peak rarefactional focal pressure-PRFP), with and without cavitation nuclei embedded within the agar at a concentration of 0.05% w/v. The second, porcine skin was insonated at 1.00 and 1.40 MPa PRFP. In both models, fluorescence measurements were used to determine penetration depth and concentration of delivered insulin. Results show that in agar gel, both insulin penetration depth and concentration only increased significantly in the presence of inertial cavitation, with up to a 40% enhancement. In porcine skin the amount of fluorescent insulin was higher in the epidermis of those samples that were exposed to ultrasound compared to the control samples, but there was no significant increase in penetration distance. The results underline the importance of instigating and monitoring inertial cavitation during transdermal insulin delivery.

Authors : Feiszthuber Helga, Bhatnagar Sunali, Gyöngy Miklós, Coussios Constantin-C,



(5) Evaluation of commercial ELISA and RIA for measuring porcine C-peptide: implications for research.[TOP]

Pubmed ID :25291621
Publication Date : //
Pre-clinical demonstration of porcine islet graft function is necessary to support the clinical transplantation of pig islets. C-peptide concentration is an especially useful marker of insulin secretion, because its measurement is not confounded by the presence of exogenous insulin. To measure porcine C-peptide (PCP), researchers in the field exclusively used the Millipore (previously Linco Research) radioimmunoassay (RIA) until 2011, when Mercodia released an alternative enzyme-linked immunosorbent assay (ELISA). (At the end of 2013, the Millipore RIA was withdrawn from the market for commercial reasons.) In our current study, to directly compare these two assays, we performed validation studies on each. We also performed interlaboratory comparisons. Then, to determine the level of agreement between the assays, we analyzed the porcine serum C-peptide concentration measurement results obtained from each assay.

Authors : Graham Melanie L, Gresch Sarah C, Hardy Sharon K, Mutch Lucas A, Janecek Jody L, Hegstad-Davies Rebecca L,



(6) Subnormothermic preservation in somah: a novel approach for enhanced functional resuscitation of donor hearts for transplant.[TOP]

Pubmed ID :25154901
Publication Date : //
Organ preservation at 4°C results in temporally irreversible injury to cellular structure and function. This study was designed to evaluate the possibility of storing hearts at ambient temperatures in novel organ preservation solution Somah to prevent damage and preserve optimum function by maintaining cellular energy over the temperature range of storage. Porcine hearts were stored in Celsior at 4°C and Somah at 4°C, 13°C and 21°C for 5 h thereafter reperfused and reanimated in vitro for 3 h. Heart weights, histopathology, ultrastructure and 2-dimensional echocardiography (2D-Echo) assessments showed preservation of structure in Somah groups. Tissue high-energy phosphate levels in Somah groups after storage were significantly greater than the Celsior hearts (p < 0.05) and highest in the 21°C Somah hearts. Upon reperfusion, myocardial O2 consumption and lactate levels quickly achieved steady state in 21°C hearts, but were delayed in Somah 4/13°C groups and severely depressed in the Celsior group. Inotrope and electroconversion requirements were inversely related to storage temperature. In vitro 2D Echo demonstrated a discordantly attenuated function in the Celsior group, moderate functionality in 4°C Somah group and superior reestablishment of performance in the Somah higher temperature groups. Hearts stored in Somah at 21°C were metabolically and functionally superior to any other groups.

Authors : Lowalekar S K, Cao H, Lu X-G, Treanor P R, Thatte H S,



(7) Aggravation of cold-induced injury in Vero-B4 cells by RPMI 1640 medium - identification of the responsible medium components.[TOP]

Pubmed ID :23046946
Publication Date : //
In modern biotechnology, there is a need for pausing cell lines by cold storage to adapt large-scale cell cultures to the variable demand for their products. We compared various cell culture media/solutions for cold storage of Vero-B4 kidney cells, a cell line widely used in biotechnology.

Authors : Pless-Petig Gesine, Metzenmacher Martin, Türk Tobias R, Rauen Ursula,



(8) Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum.[TOP]

Pubmed ID :23013186
Publication Date : //
To evaluate a human radioimmunoassay (RIA) and equine and high-range porcine (hrp) species-specific ELISAs for the measurement of high serum insulin concentrations in ponies.

Authors : Borer-Weir Kate E, Bailey Simon R, Menzies-Gow Nicola J, Harris Patricia A, Elliott Jonathan,



(9) Species incompatibilities in the pig-to-macaque islet xenotransplant model affect transplant outcome: a comparison with allotransplantation.[TOP]

Pubmed ID :22168140
Publication Date : //
Porcine islet transplantation into diabetic non-human primates is considered most relevant in translational research supporting a clinical application. Most studies have focused on immunosuppressive protocols, while metabolic aspects have mainly been utilized in graft monitoring. We evaluated data from our group regarding human and non-human primate (NHP) allotransplantation and pig-to-NHP xenotransplantation to identify incompatibilities in metabolic factors and their consequences for transplant outcomes.

Authors : Graham Melanie L, Bellin Melena D, Papas Klearchos K, Hering Bernhard J, Schuurman Henk-Jan,



(10) Cardiovascular and metabolic effects of high-dose insulin in a porcine septic shock model.[TOP]

Pubmed ID :20370783
Publication Date : //
High-dose insulin (HDI) has inotropic and vasodilatory properties in various clinical conditions associated with myocardial depression. The authors hypothesized that HDI will improve the myocardial depression produced by severe septic shock and have beneficial effects on metabolic parameters. In an animal model of severe septic shock, this study compared the effects of HDI treatment to normal saline (NS) resuscitation alone.

Authors : Holger Joel S, Dries David J, Barringer Kelly W, Peake Benjamin J, Flottemesch Thomas J, Marini John J,