„I worked with Nikolay for 2 years at EMBL Hamburg. I learnt a lot from him on protein expression in mammalian cells and this is far beyond a protocol execution. He was always supportive and open and shared his techniques in lab without hesitation. On a personal level, Nikolay is very communicative, reliable, organized, supportive, and optimistic. It was a real pleasure to both work and chat with him. Last but not least, I was amazed by Nikolay‘s ability to always learn new things. He is curious about method developments and optimization and finds solutions to all challenges on his way. “
Nikolay Dobrev
Göttingen, Niedersachsen, Deutschland
6826 Follower:innen
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With over 12 years of experience in protein expression, purification, and biophysical…
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Dear Network, Would you like to join our state-of-the-art #MS #characterization lab at #Symphogen? We are working with super interesting and…
Dear Network, Would you like to join our state-of-the-art #MS #characterization lab at #Symphogen? We are working with super interesting and…
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🎯 Protein Production Tips – Part 21: 𝐖𝐏𝐑𝐄 – 𝐓𝐡𝐞 𝐔𝐧𝐬𝐮𝐧𝐠 𝐇𝐞𝐫𝐨 𝐓𝐡𝐚𝐭 𝐒𝐮𝐩𝐞𝐫𝐜𝐡𝐚𝐫𝐠𝐞𝐬 𝐘𝐨𝐮𝐫 𝐏𝐫𝐨𝐭𝐞𝐢𝐧…
🎯 Protein Production Tips – Part 21: 𝐖𝐏𝐑𝐄 – 𝐓𝐡𝐞 𝐔𝐧𝐬𝐮𝐧𝐠 𝐇𝐞𝐫𝐨 𝐓𝐡𝐚𝐭 𝐒𝐮𝐩𝐞𝐫𝐜𝐡𝐚𝐫𝐠𝐞𝐬 𝐘𝐨𝐮𝐫 𝐏𝐫𝐨𝐭𝐞𝐢𝐧…
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🎯 Protein Production Tips – Part 21: 𝐖𝐏𝐑𝐄 – 𝐓𝐡𝐞 𝐔𝐧𝐬𝐮𝐧𝐠 𝐇𝐞𝐫𝐨 𝐓𝐡𝐚𝐭 𝐒𝐮𝐩𝐞𝐫𝐜𝐡𝐚𝐫𝐠𝐞𝐬 𝐘𝐨𝐮𝐫 𝐏𝐫𝐨𝐭𝐞𝐢𝐧…
🎯 Protein Production Tips – Part 21: 𝐖𝐏𝐑𝐄 – 𝐓𝐡𝐞 𝐔𝐧𝐬𝐮𝐧𝐠 𝐇𝐞𝐫𝐨 𝐓𝐡𝐚𝐭 𝐒𝐮𝐩𝐞𝐫𝐜𝐡𝐚𝐫𝐠𝐞𝐬 𝐘𝐨𝐮𝐫 𝐏𝐫𝐨𝐭𝐞𝐢𝐧…
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McQ – An open-source multiplexed SARS-CoV-2 quantification platform
https://www.medrxiv.org/
McQ is a SARS-CoV-2 quantification assay that couples early-stage barcoding with high-throughput sequencing to enable multiplexed processing of thousands of samples. McQ is based on homemade enzymes to enable low-cost testing of large sample pools, circumventing supply chain shortages.
Implementation of cost-efficient high-throughput methods for detection of RNA viruses such as SARS-CoV-2 is a potent strategy to curb ongoing and future pandemics. Here we describe Multiplexed SARS-CoV-2…McQ is a SARS-CoV-2 quantification assay that couples early-stage barcoding with high-throughput sequencing to enable multiplexed processing of thousands of samples. McQ is based on homemade enzymes to enable low-cost testing of large sample pools, circumventing supply chain shortages.
Implementation of cost-efficient high-throughput methods for detection of RNA viruses such as SARS-CoV-2 is a potent strategy to curb ongoing and future pandemics. Here we describe Multiplexed SARS-CoV-2 Quantification platform (McQ), an in-expensive scalable framework for SARS-CoV-2 quantification in saliva samples. McQ is based on the parallel sequencing of barcoded amplicons generated from SARS- CoV-2 genomic RNA. McQ uses indexed, target-specific reverse transcription (RT) to generate barcoded cDNA for amplifying viral- and human-specific regions. The barcoding system enables early sample pooling to reduce hands-on time and makes the ap-proach scalable to thousands of samples per sequencing run. Robust and accurate quantification of viral load is achieved by measuring the abundance of Unique Molecular Identifiers (UMIs) introduced during reverse transcription. The use of homemade reverse transcriptase and polymerase enzymes and non-proprietary buffers reduces RNA to library reagent costs to 92 cents/sample and circumvents potential supply chain short-ages. We demonstrate the ability of McQ to robustly quantify various levels of viral RNA in 838 clinical samples and accu-rately diagnose positive and negative control samples in a test-ing workflow entailing self-sampling and automated RNA ex-traction from saliva. The implementation of McQ is modular, scalable and could be extended to other pathogenic targets in future.Andere Autor:innenVeröffentlichung anzeigen -
Fast and inexpensive whole genome sequencing library preparation from intact yeast cells
G3
Through the increase in the capacity of sequencing machines massively parallel sequencing of thousands of samples in a single run is now possible. With the improved throughput and resulting drop in the price of sequencing, the cost and time for preparation of sequencing libraries have become the major bottleneck in large-scale experiments. Methods using a hyperactive variant of the Tn5 transposase efficiently generate libraries starting from cDNA or genomic DNA in a few hours and are highly…
Through the increase in the capacity of sequencing machines massively parallel sequencing of thousands of samples in a single run is now possible. With the improved throughput and resulting drop in the price of sequencing, the cost and time for preparation of sequencing libraries have become the major bottleneck in large-scale experiments. Methods using a hyperactive variant of the Tn5 transposase efficiently generate libraries starting from cDNA or genomic DNA in a few hours and are highly scalable. For genome sequencing, however, the time and effort spent on genomic DNA isolation limits the practicability of sequencing large numbers of samples. Here, we describe a highly scalable method for preparing high quality whole-genome sequencing libraries directly from yeast cultures in less than three hours at 34 cents per sample. We skip the rate-limiting step of genomic DNA extraction by directly tagmenting yeast spheroplasts and add a nucleosome release step prior to enrichment PCR to improve the evenness of genomic coverage. Resulting libraries do not show any GC-bias and are comparable in quality to libraries processed from genomic DNA with a commercially available Tn5-based kit. We use our protocol to investigate CRISPR/Cas9 on- and off-target edits and reliably detect edited variants and shared polymorphisms between strains. Our protocol enables rapid preparation of unbiased and high-quality, sequencing-ready indexed libraries for hundreds of yeast strains in a single day at a low price. By adjusting individual steps of our workflow we expect that our protocol can be adapted to other organisms.
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Selection, biophysical and structural analysis of synthetic nanobodies that effectively neutralize SARS-CoV-2
Nat Commun
The coronavirus SARS-CoV-2 is the cause of the ongoing COVID-19 pandemic. Therapeutic neutralizing antibodies constitute a key short-to-medium term approach to tackle COVID-19. However, traditional antibody production is hampered by long development times and costly production. Here, we report the rapid isolation and characterization of nanobodies from a synthetic library, known as sybodies (Sb), that target the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Several binders with…
The coronavirus SARS-CoV-2 is the cause of the ongoing COVID-19 pandemic. Therapeutic neutralizing antibodies constitute a key short-to-medium term approach to tackle COVID-19. However, traditional antibody production is hampered by long development times and costly production. Here, we report the rapid isolation and characterization of nanobodies from a synthetic library, known as sybodies (Sb), that target the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Several binders with low nanomolar affinities and efficient neutralization activity were identified of which Sb23 displayed high affinity and neutralized pseudovirus with an IC50 of 0.6 µg/ml. A cryo-EM structure of the spike bound to Sb23 showed that Sb23 binds competitively in the ACE2 binding site. Furthermore, the cryo-EM reconstruction revealed an unusual conformation of the spike where two RBDs are in the ‘up’ ACE2-binding conformation. The combined approach represents an alternative, fast workflow to select binders with neutralizing activity against newly emerging viruses.
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Transient RNA-DNA Hybrids Are Required for Efficient Double-Strand Break Repair
Cell
RNA-DNA hybrids are a major internal cause of DNA damage within cells, and their degradation by RNase H enzymes is important for maintaining genomic stability. Here, we identified an unexpected role for RNA-DNA hybrids and RNase H enzymes in DNA repair. Using a site-specific DNA double-strand break (DSB) system in Schizosaccharomyces pombe, we showed that RNA-DNA hybrids form as part of the homologous-recombination (HR)-mediated DSB repair process and that RNase H enzymes are essential for…
RNA-DNA hybrids are a major internal cause of DNA damage within cells, and their degradation by RNase H enzymes is important for maintaining genomic stability. Here, we identified an unexpected role for RNA-DNA hybrids and RNase H enzymes in DNA repair. Using a site-specific DNA double-strand break (DSB) system in Schizosaccharomyces pombe, we showed that RNA-DNA hybrids form as part of the homologous-recombination (HR)-mediated DSB repair process and that RNase H enzymes are essential for their degradation and efficient completion of DNA repair. Deleting RNase H stabilizes RNA-DNA hybrids around DSB sites and strongly impairs recruitment of the ssDNA-binding RPA complex. In contrast, overexpressing RNase H1 destabilizes these hybrids, leading to excessive strand resection and RPA recruitment and to severe loss of repeat regions around DSBs. Our study challenges the existing model of HR-mediated DSB repair and reveals a surprising role for RNA-DNA hybrids in maintaining genomic stability.
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Differentially Secreted Proteins of Antarctic and Mesophilic Strains of Synechocystis Salina and Chlorella Vulgaris after UV-B and Temperature Stress Treatment
Biotechnology & Biotechnological Equipment
Exponential-phase cell cultures from Antarctic and mesophilic strains of the cyanobacterium Synechocystis salina and the unicellular green alga Chlorella vulgaris were subjected to UV-B and temperature stress and investigated for alterations in the spectrum of secreted proteins. In addition to constitutively secreted proteins, strain-specific stress proteins were detected on silver-stained gels. Extracellular protein patterns within a given strain, as well as between strains after exposure to…
Exponential-phase cell cultures from Antarctic and mesophilic strains of the cyanobacterium Synechocystis salina and the unicellular green alga Chlorella vulgaris were subjected to UV-B and temperature stress and investigated for alterations in the spectrum of secreted proteins. In addition to constitutively secreted proteins, strain-specific stress proteins were detected on silver-stained gels. Extracellular protein patterns within a given strain, as well as between strains after exposure to each UV-B and temperature stress were notably different.
Andere Autor:innenVeröffentlichung anzeigen
Auszeichnungen/Preise
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Attending Hackathon {Life Science} meets IT in MAFINEX, Mannheim
Cubex41, MAFINEX Gründerverbund, Heidelberg Startup Partners e.V. and Heidelberg University
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Part of winning team for Patient-Centered Innovation during Hackathon {Life Science} meets IT
EIT Health
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DAAD PhD fellowship
DAAD
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Diploma Winner of the charter class of 2009/2010 speciality Molecular Biology
Faculty of Biology, Sofia University"St. Kliment Ohridski"
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Honorary medal "National Diploma"
Ministry of Education and Science, Bulgaria
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Diploma for participation in the XVII International Biology Olympiad held in Rio Cuarto on 916 July 2006, Argentina
International Biology Olympiad Committee
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Statuette Winner of the charter class of 2006
Secondary School of Natural Sciences and Mathematics
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Diploma of Laureate (3rd place) in the XXXI National Biology Olympiad
Ministry of Education and Science, Bulgaria
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Diploma for participation in the XXX National Biology Olympiad
Ministry of Education and Science, Bulgaria
Sprachen
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Bulgarian
Muttersprache oder zweisprachig
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English
Fließend
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German
Grundkenntnisse
Erhaltene Empfehlungen
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LinkedIn Mitglied
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🎯 Protein Production Tips – Part 20: 𝐃𝐢𝐯𝐢𝐝𝐞 𝐚𝐧𝐝 𝐂𝐨𝐧𝐪𝐮𝐞𝐫 – 𝐓𝐡𝐞 𝐌𝐚𝐠𝐢𝐜 𝐨𝐟 𝐒𝐞𝐥𝐟-𝐂𝐥𝐞𝐚𝐯𝐢𝐧𝐠 𝐏𝐞𝐩𝐭𝐢𝐝𝐞𝐬 (𝐏2𝐀,…
🎯 Protein Production Tips – Part 20: 𝐃𝐢𝐯𝐢𝐝𝐞 𝐚𝐧𝐝 𝐂𝐨𝐧𝐪𝐮𝐞𝐫 – 𝐓𝐡𝐞 𝐌𝐚𝐠𝐢𝐜 𝐨𝐟 𝐒𝐞𝐥𝐟-𝐂𝐥𝐞𝐚𝐯𝐢𝐧𝐠 𝐏𝐞𝐩𝐭𝐢𝐝𝐞𝐬 (𝐏2𝐀,…
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🎯 Protein Production Tips – Part 20: 𝐃𝐢𝐯𝐢𝐝𝐞 𝐚𝐧𝐝 𝐂𝐨𝐧𝐪𝐮𝐞𝐫 – 𝐓𝐡𝐞 𝐌𝐚𝐠𝐢𝐜 𝐨𝐟 𝐒𝐞𝐥𝐟-𝐂𝐥𝐞𝐚𝐯𝐢𝐧𝐠 𝐏𝐞𝐩𝐭𝐢𝐝𝐞𝐬 (𝐏2𝐀,…
🎯 Protein Production Tips – Part 20: 𝐃𝐢𝐯𝐢𝐝𝐞 𝐚𝐧𝐝 𝐂𝐨𝐧𝐪𝐮𝐞𝐫 – 𝐓𝐡𝐞 𝐌𝐚𝐠𝐢𝐜 𝐨𝐟 𝐒𝐞𝐥𝐟-𝐂𝐥𝐞𝐚𝐯𝐢𝐧𝐠 𝐏𝐞𝐩𝐭𝐢𝐝𝐞𝐬 (𝐏2𝐀,…
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Munich Lab Automation Community Gewinnspiel!!!
Munich Lab Automation Community Gewinnspiel!!!
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🎉 A Year of Growth and New Beginnings 🚀 As the year comes to a close, I find myself reflecting on an incredible journey at IBA Lifesciences. Being…
🎉 A Year of Growth and New Beginnings 🚀 As the year comes to a close, I find myself reflecting on an incredible journey at IBA Lifesciences. Being…
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Hey, München! 👋 On January 10th, I’ll be giving a talk at the Center for NanoScience, LMU Munich on how to deliver therapeutic RNAs, major…
Hey, München! 👋 On January 10th, I’ll be giving a talk at the Center for NanoScience, LMU Munich on how to deliver therapeutic RNAs, major…
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