Name: generation of recombinant arenavirus for vaccine development in fda-approved vero cells
Uploaded: 2013-08-01T13:00:00
Description: Watch this Scientific Journal Video about Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells at JoVE.com

Vi takker tidligere og nuværende medlemmer i JCT og LM-S laboratorier til udvikling og forbedring af arenavirus teknikker til omvendt genetik og plasmider. Vi takker også Snezhana Dimitrova for teknisk support. Det monoklonale antistof rettet mod JUNV NP (SA02-BG12) blev opnået fra BEI Resources (NIAID Biodefense og Emerging Infektioner Research Resources Repository). BYHC blev støttet af Grant Number GM068411 fra Institutional Ruth L. Kirschstein National Research Service Award. EO-R er en Fullbright-Conicyt (BIO 2008) og et Rochester Vaccine Fellowship (2012) modtager. EO-R nuværende støtte ydes af en postdoc Fellowship for Mangfoldighed og Academic Excellence fra Kontoret for Faculty Development &amp; Mangfoldighed på University of Rochester. Forskning i LM-S laboratorium er finansieret af NIH tilskud RO1 AI077719, R21NS075611-01, R03AI099681-01A1, NIAID Centers of Excellence for Influenza Forskning og overvågning (HHSN266200700008C), ogThe University of Rochester Center for Biodefense Immun Modeling (HHSN272201000055C). Forskning på JCT Laboratoriet blev støttet af tilskud RO1 AI047140, RO1 AI077719 og RO1 AI079665 fra NIH.

1.. Arenavirus Rescue Transfektion Vores rednings-system var baseret på anvendelsen af ​​både polymerase II protein ekspressionsplasmider der koder for nukleoprotein (NP) og RNA-afhængige RNA-polymerase-(L), de virale trans-virkende faktorer, der er nødvendige for RNA-replikation og genekspression af arenavirus genomet 7, 19, og plasmider stand til at styre intracellulær syntese, via cellulært RNA-polymerase I (pol-I), af S-og L antigenom RNA-art 7. I vores unde...

<ol>
	<li>Albarino, C. G., Bird, B. H., Chakrabarti, A. K., Dodd, K. A., Flint, M., Bergeron, E., White, D. M., Nichol, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+major+determinant+of+attenuation+in+mice+of+the+Candid1+vaccine+for+Argentine+hemorrhagic+fever+is+located+in+the+G2+glycoprotein+transmembrane+domain.">The major determinant of attenuation in mice of the Candid1 vaccine for Argentine hemorrhagic fever is located in the G2 glycoprotein transmembrane domain.</a> Journal of Virology. 85, 10404-10408 (2011).</li><li>Barton, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphocytic+choriomeningitis+virus:+a+neglected+central+nervous+system+pathogen.">Lymphocytic choriomeningitis virus: a neglected central nervous system pathogen.</a> Clinical Infectious Diseases: an official publication of the Infectious Diseases Society of America. 22, 197 (1996).</li><li>Buchmeier, M. J., Peter, C. J., de la Torre, J. C., Fields, B. N., Knipe, D. M., Howley, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arenaviridae:+The+viruses+and+their+replication.">Arenaviridae: The viruses and their replication.</a> Fields' Virology. 2, 179201827 (2007).</li><li>Emonet, S. E., Urata, S., de la Torre, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arenavirus+reverse+genetics:+new+approaches+for+the+investigation+of+arenavirus+biology+and+development+of+antiviral+strategies.">Arenavirus reverse genetics: new approaches for the investigation of arenavirus biology and development of antiviral strategies.</a> Virology. 411, 416-425 (2011).</li><li>Emonet, S. F., Garidou, L., McGavern, D. B., de la Torre, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+recombinant+lymphocytic+choriomeningitis+viruses+with+trisegmented+genomes+stably+expressing+two+additional+genes+of+interest.">Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest.</a> Proceedings of the National Academy of Sciences of the United States of America. 106, 3473-3478 (2009).</li><li>Fischer, S. A., Graham, M. B., Kuehnert, M. J., Kotton, C. N., Srinivasan, A., Marty, F. M., Comer, J. A., Guarner, J., Paddock, C. D., DeMeo, D. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transmission+of+lymphocytic+choriomeningitis+virus+by+organ+transplantation.">Transmission of lymphocytic choriomeningitis virus by organ transplantation.</a> The New England Journal of Medicine. 354, 2235-2249 (2006).</li><li>Flatz, L., Bergthaler, A., de la Torre, J. C., Pinschewer, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recovery+of+an+arenavirus+entirely+from+RNA+polymerase+I/II-driven+cDNA.">Recovery of an arenavirus entirely from RNA polymerase I/II-driven cDNA.</a> Proceedings of the National Academy of Sciences of the United States of America. 103, 4663-4668 (2006).</li><li>Gunther, S., Lenz, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lassa+virus.">Lassa virus.</a> Critical reviews in clinical laboratory sciences. 41, 339-390 (2004).</li><li>Harrison, L. H., Halsey, N. A., McKee, K. T., Peters, C. J., Barrera Oro, J. G., Briggiler, A. M., Feuillade, M. R., Maiztegui, J. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+case+definitions+for+Argentine+hemorrhagic+fever.">Clinical case definitions for Argentine hemorrhagic fever.</a> Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America. 28, 1091-1094 (1999).</li><li>Isaacson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+hemorrhagic+fever+hazards+for+travelers+in+Africa.">Viral hemorrhagic fever hazards for travelers in Africa.</a> Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America. 33, 1707-1712 (2001).</li><li>Kerber, R., Rieger, T., Busch, C., Flatz, L., Pinschewer, D. D., Kummerer, B. M., Gunther, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-species+analysis+of+the+replication+complex+of+Old+World+arenaviruses+reveals+two+nucleoprotein+sites+involved+in+L+protein+function.">Cross-species analysis of the replication complex of Old World arenaviruses reveals two nucleoprotein sites involved in L protein function.</a> Journal of Virology. 85, 12518-12528 (2011).</li><li>Lee, K. J., Novella, I. S., Teng, M. N., Oldstone, M. B., de La Torre, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NP+and+L+proteins+of+lymphocytic+choriomeningitis+virus+(LCMV)+are+sufficient+for+efficient+transcription+and+replication+of+LCMV+genomic+RNA+analogs.">NP and L proteins of lymphocytic choriomeningitis virus (LCMV) are sufficient for efficient transcription and replication of LCMV genomic RNA analogs.</a> Journal of Virology. 74, 3470-3477 (2000).</li><li>Mets, M. B., Barton, L. L., Khan, A. S., Ksiazek, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphocytic+choriomeningitis+virus:+an+underdiagnosed+cause+of+congenital+chorioretinitis.">Lymphocytic choriomeningitis virus: an underdiagnosed cause of congenital chorioretinitis.</a> American Journal of Ophthalmology. 130, 209-215 (2000).</li><li>Murakami, S., Horimoto, T., Yamada, S., Kakugawa, S., Goto, H., Kawaoka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+canine+RNA+polymerase+I-driven+reverse+genetics+for+influenza+A+virus:+its+application+for+H5N1+vaccine+production.">Establishment of canine RNA polymerase I-driven reverse genetics for influenza A virus: its application for H5N1 vaccine production.</a> Journal of Virology. 82, 1605-1609 (2008).</li><li>Palacios, G., Druce, J., Du, L., Tran, T., Birch, C., Briese, T., Conlan, S., Quan, P. L., Hui, J., Marshall, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+arenavirus+in+a+cluster+of+fatal+transplant-associated+diseases.">A new arenavirus in a cluster of fatal transplant-associated diseases.</a> The New England Journal of Medicine. 358, 991-998 (2008).</li><li>Rodrigo, W. W., de la Torre, J. C., Martinez-Sobrido, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+single-cycle+infectious+lymphocytic+choriomeningitis+virus+to+study+hemorrhagic+fever+arenaviruses.">Use of single-cycle infectious lymphocytic choriomeningitis virus to study hemorrhagic fever arenaviruses.</a> Journal of Virology. 85, 1684-1695 (2011).</li><li>Rojek, J. M., Kunz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+entry+by+human+pathogenic+arenaviruses.">Cell entry by human pathogenic arenaviruses.</a> Cellular Microbiology. 10, 828-835 (2008).</li><li>Rojek, J. M., Sanchez, A. B., Nguyen, N. T., de la Torre, J. C., Kunz, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+mechanisms+of+cell+entry+by+human-pathogenic+Old+World+and+New+World+arenaviruses.">Different mechanisms of cell entry by human-pathogenic Old World and New World arenaviruses.</a> Journal of Virology. 82, 7677-7687 (2008).</li><li>Sanchez, A. B., de la Torre, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescue+of+the+prototypic+Arenavirus+LCMV+entirely+from+plasmid.">Rescue of the prototypic Arenavirus LCMV entirely from plasmid.</a> Virology. 350, 370-380 (2006).</li></ol>

Generering af rekombinante arenaviruses hjælp plasmid-baserede teknikker til omvendt genetik er blevet en udbredt metode til undersøgelse af mange forskellige facetter af arenavirus biologi. Her har vi dokumentere en afgørende forbedring af det nuværende system ved at udføre arenavirus redninger i Vero-celler, der giver mulighed for den potentielle produktion af FDA-godkendte vaccinekandidater mod arenaviruses og vaccine vektorer mod andre smitsomme sygdomme. De eksperimentelle involver...

Arenaviruses er indkapslede vira med bisegmented, negativ-strenget RNA-genom 3, som hører til Arenaviridae familien. Den arenavirus genomet koder fire proteiner i en ambisense mode fra to separate virale segmenter 3. Den store (L) segment koder for RNA-afhængige RNA-polymerase (L) og den lille RING (Virkelig interessant nyt gen) finger protein (Z), der tjener som den vigtigste drivkraft for virus spirende. Den lille (S) segment koder for virale nukleoprotein (NP) og overfladen glycoprotein (GP) (Figur 1). Adskillige medlemmer af Arenaviridae familien er ansvarlige for dødelig hæmoragisk feber (HF) i mennesker 3. Principielle bekymringer er Lassa virus (LASV), og Junín virus (JUNV), der er kendt for at forårsage høj dødelighed hos indlagte patienter 8, 9. Selv om disse vira er endemiske for Vestafrika og landdistrikter Argentina henholdsvisDer er stigende bekymring for import af LASV og JUNV til ikke-endemiske områder på grund af øget rejse 10. Derudover, selvom ikke normalt er forbundet med alvorlig sygdom hos mennesker, er prototypisk arenavirus lymfatisk choriomeningitisvirus (LCMV) betragtes som en forsømt patogen som der har været tilfælde af dødelig infektion i immunsvækkede patienter 6, 15 og er ansvarlig for medfødte fosterskader og spontane aborter hos gravide kvinder 2, 13. I øjeblikket er der ingen amerikanske FDA-godkendte vacciner mod arenaviruses og behandling er begrænset til nukleosidanalogen ribavirin, som kun er delvist effektiv og ofte forbundet med betydelige bivirkninger. Indførelsen af plasmid-baserede revers genetik 4 og generering af rekombinante arenaviruses 7, 19 har gjort store fremskridt inden for arenavirus forskning. I øjeblikket er gnaverceller (såsom BHK-21) anvendes til generelttion af rekombinante arenaviruses grund af artsspecifikke murine RNA polymerase I (pol-I) promotor, der styrer den første transkription af S og L segmenter. Men virus redning i BHK-21-celler er ikke en godkendt metode til generering af rekombinante arenaviruses som potentielle vaccine frø kandidater. Her har vi dokumentere brugen af ​​menneskelige pol-I promoter til effektiv redning af prototypisk gamle verden (OW) LCMV og den nye verden (NW) JUNV Candid # 1 stamme i Vero-celler. Ved hjælp af en lignende metode, men vi genererede rekombinant trisegmented LCMV (r3LCMV), og Candid # 1 (r3Candid # 1) arenaviruses, der indeholder yderligere to fremmede gener indkodet i to forskellige S RNA segmenter 5. Dette nye system ikke blot følger en meget reproducerbar og enkel protokol, men umiddelbart kan gennemføres i dannelsen af ​​rekombinante arenaviruses som potentiel vaccine eller vaccine vektor frø.

<table border="1">
	<tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Material and methods</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>
 Cell lines 
Vero E6 (African green monkey kidney epithelial cells) are maintained in a 37 &deg;C incubator with 5 % CO2 in DMEM 10 % FBS 1 % PS. Cells are available from the American Type Culture Collection (ATCC, catalogue number CRL-1586). 
 
Plasmids 
All plasmids, with the exception of hpol-I L, can be grown at 37 &deg;C, for 16-18 hr. We recommend that cultures of the hpol-I L be grown at 30 &deg;C for 24 hr. Plasmids are prepared using a plasmid maxi kit (EZNA Fastfilter Plasmid Maxi Kit, Omega Bio-tek) following the manufacturer's recommendations and stored at -20 &deg;C. The concentration of the purified DNA plasmid is determined by spectrophotometry at 260 nm, with purity being estimated using the 260:280 nm ratio. Preparations with 1.8-2.0 260:280 nm ratios are considered appropriate for virus rescue purposes. Additionally, plasmid concentration and purity should be confirmed with agarose gel electrophoresis. 
 
Viruses 
The described protocol for rescuing rLCMV (Armstrong 53b) and rCandid#1 can be executed under biosafety level (BSL) 2 conditions. Contaminated material, including TCS and cells should be sterilized before disposal. Rescue of other arenavirus may require higher BSL facilities so proper safety/security measures must be followed. 
 
Tissue culture media and solutions 
DMEM 10 %FBS 1 %PS: 445 ml Dulbecco's modified Eagle's medium (DMEM), 50 ml of Fetal Bovine Serum (FBS), and 5 ml of 100X Penicillin/Streptomycin (PS). Store at 4 &deg;C. This media will be used for maintenance of Vero cells. 
 
Infectious Media: 2-to-1 mixture of OptiMEM and DMEM 10 %FBS 1 %PS. Store at 4 &deg;C. This media will be used during viral infections. 
 
10X Phosphate buffered saline (PBS): 80 g of NaCl, 2 g of KCl, 11.5 g of Na2HPO4.7H2O, 2 g of KH2PO4. Add ddH2O up to 1 liter. Adjust pH to 7.3. Sterilize by autoclave. Store at room temperature. 
1X PBS: Dilute 10X PBS 1:10 with ddH2O. Sterilize by autoclave and store at room temperature. 
2.5 % BSA: 2.5 g of BSA in 97.5 ml of 1X PBS. Store at 4 &deg;C. This is used as a blocking solution for IFA.
</td>
 </tr>
 </tbody>
 </table>

generation of recombinant arenavirus for vaccine development in fda-approved vero cells

Udviklingen og implementeringen af arenavirus omvendt genetik repræsenterer et betydeligt gennembrud i arenavirus banehalvdel 4. Brugen af ​​cellebaserede arenavirus minigenomplasmiderne systemer sammen med evnen til at generere rekombinante infektiøse arenaviruses med forudbestemte mutationer i deres genomer har lettet undersøgelse af bidraget af virale determinanter til de forskellige trin i arenavirus livscyklus, samt virus-host interaktioner og mekanismer arenavirus patogenese 1, 3, 11. Desuden har udviklingen af trisegmented arenaviruses tilladt brugen af arenavirus genom at udtrykke yderligere fremmede gener af interesse og dermed åbne muligheden for arenavirus-baserede vaccine vektor applikationer 5. Ligeledes er udviklingen af ​​single-cyklus smitsomme arenaviruses stand til at udtrykke reportergener giver et nyt eksperimentelt værktøj til at forbedre sikkerheden for forskning, som involverer highly patogene humane arenaviruses 16.. Den generation af rekombinante arenaviruses hjælp plasmid-baserede teknikker til omvendt genetik har hidtil påberåbt brugen af gnaver cellelinjer 7,19, hvilket skaber nogle barrierer for udviklingen af Food and Drug Administration (FDA)-godkendt vaccine eller vaccine vektorer. For at overvinde denne hindring, vi beskriver her en effektiv generation af rekombinante arenaviruses i FDA-godkendte Vero celler.

Vellykket redning af en rekombinant vildtype arenavirus vil blive bekræftet ved tilstedeværelsen af virale antigener ved hjælp af IFA (figur 5). I tilfælde af rekombinante trisegmented vira, kan vellykket viral redning vurderes ved at observere GFP-ekspression ved fluorescensmikroskopi (Figur 6A). Vellykket redning vil blive yderligere bekræftet ved at vurdere Gluc udtryk (figur 6B). Repræsentative resultater illustrerer vellykket redning af vildtype og trisegment...

Watch this Scientific Journal Video about Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells at JoVE.com

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

Fremstilling af rekombinant arenavirus for Vaccine Development i FDA-godkendte Vero celler

Redning af rekombinante arenaviruses fra klonede cDNA&#39;er, en tilgang kaldet reverse genetik, giver forskerne at undersøge betydningen af ​​specifikke virale genprodukter, samt bidrag af deres forskellige specifikke domæner og restprodukter til mange forskellige aspekter af biologi arenavirus . Ligeledes omvendt genetik teknikker i FDA-godkendte cellelinier (Vero) for udvikling af vacciner giver nye muligheder til generering af effektive og sikre vacciner til bekæmpelse af humane patogene arenaviruses.

The development and implementation of arenavirus reverse genetics represents a significant breakthrough in the arenavirus field  4. The use of cell-based ...

Research

JoVE Journal

Immunology and Infection

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular ...

Generation of Recombinant Influenza Virus from Plasmid DNA

Efforts by a number of influenza research groups have been pivotal in the development and improvement of influenza A virus reverse genetics. Originally ...

Generation of a Novel Dendritic-cell Vaccine Using Melanoma and Squamous Cancer Stem Cells

Generation af en ny dendritisk-celle vaccine ved hjælp af melanom og squamous cancer stamceller

We identified cancer stem cell (CSC)-enriched populations from murine melanoma D5 syngeneic to C57BL/6 mice and the squamous cancer SCC7 syngeneic to C3H ...

Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis

Brug Klik Chemistry at måle effekten af ​​Viral infektion på Host-celle RNA-syntese

Many RNA viruses have evolved the ability to inhibit host cell transcription as a means to circumvent cellular defenses. For the study of these viruses, ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In vitro analyse af MyD88-medieret Cellular immunrespons til West Nile Virus mutantstamme Infektion

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

Utilizing the Antigen Capsid-Incorporation Strategy for the Development of Adenovirus Serotype 5-Vectored Vaccine Approaches

Udnytte den Antigen capsidet-Indbygning strategi for udvikling af adenovirusserotype 5-vektoriserede Vaccine Approaches

Adenovirus serotype 5 (Ad5) has been extensively modified with traditional transgene methods for the vaccine development. The reduced efficacies of these ...

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

Fremstilling af rekombinant humant IgG monoklonale antistoffer fra Immortaliserede Sorteret B-celler

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, ...

Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment

Meget følsom analyse for måling af arenavirus-celle Attachment

Arenaviruses are a family of enveloped RNA viruses that cause severe human disease. The first step in the arenavirus life cycle is attachment of viral ...

Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-&#945;

Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â±

Udvikling og validering af et Ultrasensitive enkelt molekyle Array Digital enzym-forbundet Immunosorbent Assay for menneskelige Interferon-α

The main aim of this protocol is to describe the development and validation of an interferon (IFN)-&#945; single molecule array digital Enzyme-Linked ...

Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses

Generation, forstærkning og titrering af rekombinant respiratorisk Syncytial virus

The use of recombinant viruses has become crucial in basic or applied virology. Reverse genetics has been proven to be an extremely powerful technology, ...