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European Journal of Microbiology and Immunology
Authors: Ingrid E. Pereira, Kyssia P. Silva, Laura M. Menegati, Aimara C. Pinheiro, Elaine A. O. Assunção, Maria De Lourdes P. Araújo, Elfadil Abass, Malcolm S. Duthie, Ulrich Steinhoff, and Henrique C. Teixeira

species but also trypanosomatids and even phylogenetically distant species that are co-endemic [ 7 ]. Several recombinant proteins have been evaluated for their diagnostic potential of human leishmaniasis. For example, the rLb6H protein, identified through

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The current method to detect antibody titre against infectious bursal disease virus (IBDV) in chickens is based on enzyme-linked immunosorbent assay (ELISA) using whole virus as coating antigen. Coating the ELISA plates requires a purified or at least semi-purified preparation of virus as antigen, which needs special skills and techniques. In this study, instead of using whole virus, recombinant protein of hexahistidine tag (His 6 tag) and VPX protein of IBDV expressed in E. coli was used as an alternative antigen to coat the ELISA plates. There was a good correlation coefficient (R 2 = 0.972) between the results of the ELISA using plates coated with monoclonal antibody against His 6 tag and those of the commercial IBDV ELISA kit. Hence, His 6 tag and VPX recombinant protein expressed in E. coli has the potential for the development of ELISA for the measurement of IBDV-specific antibody.

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Abstract  

After a formal explanation of Mayer's enthalpy balance method as applied to biological reaction rates, the history of its application is traced from Rubner's dog to accounting for the energy of muscle contraction. The introduction of microcalorimetry allowed the method generally to be used for cells in vitro and now particular emphasis can be paid to the growth of cells for the production of therapeutically-important heterologous proteins. In these systems, enthalpy balance studies contribute to defining catabolic processes, designing media, understanding the mechanisms of growth and controlling cultures using heat flux as an on-line sensor of metabolic activity.

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Acta Biologica Hungarica
Authors: Anna Tóth, Katalin Fodor, P. Blazsó, I. Cserpán, Tünde Praznovszky, V. Tubak, A. Udvardy, Gy. Hadlaczky, and R. Katona

Zhou, H., Wu, S., Joo, J., Y., Zhu, S., Han, D., W., Lin, T., Trauger, S., Bien, G., Yao, S., Zhu, Y., Siuzdak, G., Schöler, H., R., Duan, L., Ding, S. (2009) Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell 4

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. & Brierley, R. A. (1989): Methylotrophic yeast Pichia pastoris produced in high cell density fermentations with high cell yields as vehicle for recombinant protein production. Biotech. Bioengng. , 34 , 403-104. Methylotrophic

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Two indirect ELISAs for the detection of antibodies against glycoprotein E (gE) of Aujeszky's disease virus (ADV) in sera have been developed. The rec-gE-ELISA is based on theE. coli-expressed recombinant protein containing the N-terminal sequences of gE (aa 1-125) fused with the glutathione S-transferase fromSchistosoma japonicum. The affi-gE-ELISA is based on native gE, which was purified from virions by affinity chromatography. The tests were optimised and compared with each other, as well as with the recently developed blocking gE-ELISA (Morenkov et al., 1997b), with respect to specificity and sensitivity. The rec-gE-ELISA was less sensitive in detecting ADV-infected animals than the affi-gE-ELISA (sensitivity 80% and 97%, respectively), which is probably due to the lack of conformation-dependent immunodominant epitopes on the recombinant protein expressed inE. coli. The specificity of the rec-gE-ELISA and affi-gE-ELISA was rather moderate (90% and 94%, respectively) because it was necessary to set such cut-off values in the tests that provided a maximum level of sensitivity, which obviously increased the incidence of false positive reactions. Though the indirect ELISAs detect antibodies against many epitopes of gE, the blocking gE-ELISA, which detects antibodies against only one immunodominant epitope of gE, showed a better test performance (specificity 99% and sensitivity 98%). This is most probably due to rather high dilutions of the sera used in the indirect gE-ELISAs (1:30) as compared to the serum dilution in the blocking gE-ELISA (1:2). We conclude that the indirect gE-ELISAs are sufficiently specific and sensitive to distinguish ADV-infected swine from those vaccinated with gE-negative vaccine and can be useful, in particularly affi-gE-ELISA, as additional tests for the detection of antibodies to gE.

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. 384 105 116 Griffin, B. A., Adams, S. R., Tsien, R. Y. (1998) Specific covalent labeling of recombinant protein molecules inside live cells. Science

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). Purification of the recombinant Erp, HspR, LppX, MmaA4, and OmpA proteins Purification of the recombinant proteins was performed according to Okay et al. [ 15 ]. The recombinant E. coli BL21(DE3) cells with pET-28a(+) vectors carrying erp, hspR

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Monoklonális antitestek és egyéb biológiai terápiák a COVID–19 kezelésére

Monoclonal antibodies and other biologics for treatment of COVID-19

Scientia et Securitas
Author: Imre Kacskovics

Összefoglaló. A SARS-CoV-2 koronavírus által kiváltott pandémia az elmúlt mintegy 100 év legsúlyosabb közegészségügyi, gazdasági és társadalmi válságát okozza. Az emberiség soha nem látott tudással, példa nélküli sebességgel állította elő azokat a hatékony védőoltásokat, amelyek a megfelelő átoltottság mellett kontrollálhatják a COVID–19-járványt.

A SARS-CoV-2 fertőzéssel az emberiségnek meg kell tanulnia együtt élni, és mivel a vakcinák nem mindenkinek adhatók, vagy nem mindenkiben váltanak ki immunvédelmet, szükséges a jelenleginél hatékonyabb COVID–19-specifikus terápiák kifejlesztése. Több COVID–19 kezelésére kifejlesztett gyógyszerhatóanyagot is sikerrel tesztelnek, közülük is kiemelkednek a monoklonális antitestek, illetve más biológiai terápiák. Ezek egyfelől olyan gyógyszerek, amelyek a betegség korai fázisában semlegesítik a vírust, azaz megakadályozzák, hogy a sejteket megfertőzze, míg mások a már kialakult súlyos megbetegedésben csökkenthetik a gyulladás mértékét. Biologikumok közé tartozik a SARS-CoV-2-t semlegesítő hACE2-Fc fúziós fehérje is, amely a neutralizáló monoklonális antitestek hatásához hasonlítható; előnye, hogy minden mutáns ellen hatékony lehet.

Virológusok, járványügyi szakemberek szerint fel kell készülnünk arra, hogy a jelenlegihez hasonló járványok rendszeressé válnak. Ökológiai okok miatt egyre nő az állati eredetű kórokozók adaptálódása az emberhez, de nem zárhatók ki az ún. laborszökevény vírusok, sőt a bioterrorizmus veszélye sem. Mindezek hatékony kezelésére erősítenünk kell a hazai biotechnológiai kapacitásokat. A hatóanyagfejlesztésben a már kialakult hazai egyetemi-kutatóintézeti-ipari együttműködésekre számíthatunk, a gyártás során pedig a hazai korszerű biotechnológiai, gyógyszeripari kapacitásra, amelyek növelhetik az önellátásból származó biztonságot.

Summary. The SARS-CoV-2 coronavirus pandemic is causing the worst public health, economic and social crisis in the last 100 years. New and effective vaccines were developed and produced with the application of unprecedented know how and speed, which can control the COVID-19 epidemic with the right vaccination coverage.

Humanity needs to learn to live with the SARS-CoV-2 infection, and because vaccines cannot be given to everyone or cannot induce immune protection in all vaccinated individuals, it is necessary to develop more effective COVID-19-specific therapies than those presently available. Several drugs developed for the treatment of COVID-19 have been successfully tested, including monoclonal antibodies and other biological therapies. These are, on the one hand, drugs that neutralize the virus in the early stages of the disease, that is, it prevents it from infecting the cells, while others can reduce the rate of inflammation in a severe disease status. This review article provides an update about the current status of monoclonal antibodies that have been developed to treat COVID-19.

In early 2020 Eotvos Lorand University, Pecs University, Gedeon Richter Plc and ImmunoGenes Ltd formed a consortium to develop a molecular trap, the human ACE2-Fc fusion protein that binds to the spike protein of SARS-CoV-2 and inhibits its binding to the ACE2 receptors on the cell surfaces. We successfully produced this recombinant protein and proved that it neutralizes this virus using VERO E6 cells and protects animals (Syrian hamster) from serious disease when administered before infection. We have also shown that it has a long half-life due to its (IgG) Fc-region component. Based on these proof of concept data, we created mutant versions of this drug candidate that do not have catabolic activity for angiotensin II and thus would not influence blood pressure. This is important since this drug should be administered in log-fold higher concentrations than ACE2 receptors in the body in order to efficiently neutralize the virus. The virus neutralization capacity of these new versions remained intact based on in vitro virus neutralization tests. We believe that after successful animal experiments, these drug candidates can be efficiently used in COVID-19 therapy in mild or moderate disease status. As compared to the COVID-19 specific monoclonal antibodies, we believe that these recombinant, mutant hACE2-Fc drugs can be more effective than the mAbs as they effectively bind and neutralize the new variants of SARS-CoV-2 (if they are able to bind the endogenous ACE2 receptor).

According to virologists and epidemiologists, we need to be prepared for epidemics like the current one becoming more regular. Due to ecological reasons, the adaptation of animal pathogens to humans is increasing, but there are threats due to lab leak viruses and even bioterrorism. To deal with all this effectively, we need to strengthen domestic biotechnology capacities. In the development of drug substances, we can count on the already established Hungarian university-research institute-industry collaborations, which can increase the safety resulting from self-sufficiency.

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Acta Veterinaria Hungarica
Authors: Beáta Kovács, Mathilda Toussaint, E. Gruys, Ibolya Fábián, L. Szilágyi, J. Janan, and P. Rudas

489 503 Kovacs, B. M., Szilagyi, L., Janan, J. and Rudas, P. (2005): Serum amyloid A in geese; cloning and expression of recombinant protein. Amyloid 12 , 109

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