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Stabilization of the HIV-1 Envelope glycoprotein trimer (Env) in its native pre-fusion closed conformation is regarded as one of several requirements for the induction of neutralizing antibody (nAb) responses, which, in turn, will most likely be a prerequisite for the development of an efficacious preventive vaccine. Here, we systematically analyzed how the stepwise stabilization of a clade C consensus (ConC) Env immunogen impacts biochemical and biophysical protein traits such as antigenicity, thermal stability, structural integrity, and particle size distribution. The increasing degree of conformational rigidification positively correlates with favorable protein characteristics, leading to optimized homogeneity of the protein preparations, increased thermal stability, and an overall favorable binding profile of structure-dependent broadly neutralizing antibodies (bnAbs) and non-neutralizing antibodies (non-nAbs). We confirmed that increasing the structural integrity and stability of the Env trimers positively correlates with the quality of induced antibody responses by the immunogens. These and other data contribute to the selection of ConCv5 KIKO as novel Env immunogens for use within the European Union’s H2020 Research Consortium EHVA (European HIV Alliance) for further preclinical analysis and phase 1 clinical development.
Alexandra Hauser; George Carnell; Kathrin Held; Guidenn Sulbaran; Nadine Tischbierek; Lisa Rogers; Georgios Pollakis; Paul Tonks; Michael Hoelscher; Song Ding; Rogier Sanders; Christof Geldmacher; Quentin Sattentau; Winfried Weissenhorn; Jonathan Heeney; David Peterhoff; Ralf Wagner. Stepwise Conformational Stabilization of a HIV-1 Clade C Consensus Envelope Trimer Immunogen Impacts the Profile of Vaccine-Induced Antibody Responses. Vaccines 2021, 9, 750 .
AMA StyleAlexandra Hauser, George Carnell, Kathrin Held, Guidenn Sulbaran, Nadine Tischbierek, Lisa Rogers, Georgios Pollakis, Paul Tonks, Michael Hoelscher, Song Ding, Rogier Sanders, Christof Geldmacher, Quentin Sattentau, Winfried Weissenhorn, Jonathan Heeney, David Peterhoff, Ralf Wagner. Stepwise Conformational Stabilization of a HIV-1 Clade C Consensus Envelope Trimer Immunogen Impacts the Profile of Vaccine-Induced Antibody Responses. Vaccines. 2021; 9 (7):750.
Chicago/Turabian StyleAlexandra Hauser; George Carnell; Kathrin Held; Guidenn Sulbaran; Nadine Tischbierek; Lisa Rogers; Georgios Pollakis; Paul Tonks; Michael Hoelscher; Song Ding; Rogier Sanders; Christof Geldmacher; Quentin Sattentau; Winfried Weissenhorn; Jonathan Heeney; David Peterhoff; Ralf Wagner. 2021. "Stepwise Conformational Stabilization of a HIV-1 Clade C Consensus Envelope Trimer Immunogen Impacts the Profile of Vaccine-Induced Antibody Responses." Vaccines 9, no. 7: 750.
The delivery of HIV-1 envelope (Env) trimer-based immunogens on the surface of nanoparticles holds promise to promote immunogenicity with the aim of inducing a potent, durable and broad neutralizing antibody (bnAb) response. Towards that goal, we examined the covalent conjugation of Env to 100 nm and 200 nm silica nanoparticles (SiNPs) to optimize conjugation density and attachment stability. Env was redesigned to enable site-specific cysteine-mediated covalent conjugation while maintaining its structural integrity and antigenicity. Env was anchored to different sized SiNPs with a calculated spacing of 15 nm between adjacent trimers. Both particle sizes exhibited high in vitro stability over a seven-day period. After attachment, 100 nm particles showed better colloidal stability compared to 200 nm particles. Importantly, the antigenic profile of Env was not impaired by surface attachment, indicating that the quaternary structure was maintained. In vitro Env uptake by dendritic cells was significantly enhanced when Env was delivered on the surface of nanoparticles compared to soluble Env. Furthermore, multivalent Env displayed efficiently activated B cells even at Env concentrations in the low nanomolar range. In mice, antibody responses to nanoparticle-coupled Env were stronger compared to the free protein and had equivalent effects at lower doses and without adjuvant.
David Peterhoff; Stefanie Thalhauser; Jan Sobczak; Mona Mohsen; Christoph Voigt; Nicole Seifert; Patrick Neckermann; Alexandra Hauser; Song Ding; Quentin Sattentau; Martin Bachmann; Miriam Breunig; Ralf Wagner. Augmenting the Immune Response against a Stabilized HIV-1 Clade C Envelope Trimer by Silica Nanoparticle Delivery. Vaccines 2021, 9, 642 .
AMA StyleDavid Peterhoff, Stefanie Thalhauser, Jan Sobczak, Mona Mohsen, Christoph Voigt, Nicole Seifert, Patrick Neckermann, Alexandra Hauser, Song Ding, Quentin Sattentau, Martin Bachmann, Miriam Breunig, Ralf Wagner. Augmenting the Immune Response against a Stabilized HIV-1 Clade C Envelope Trimer by Silica Nanoparticle Delivery. Vaccines. 2021; 9 (6):642.
Chicago/Turabian StyleDavid Peterhoff; Stefanie Thalhauser; Jan Sobczak; Mona Mohsen; Christoph Voigt; Nicole Seifert; Patrick Neckermann; Alexandra Hauser; Song Ding; Quentin Sattentau; Martin Bachmann; Miriam Breunig; Ralf Wagner. 2021. "Augmenting the Immune Response against a Stabilized HIV-1 Clade C Envelope Trimer by Silica Nanoparticle Delivery." Vaccines 9, no. 6: 642.
SARS-CoV-2 infection fatality ratios (IFR) remain controversially discussed with implications for political measures. The German county of Tirschenreuth suffered a severe SARS-CoV-2 outbreak in spring 2020, with particularly high case fatality ratio (CFR). To estimate seroprevalence, underreported infections, and IFR for the Tirschenreuth population aged ≥14 years in June/July 2020, we conducted a population-based study including home visits for the elderly, and analyzed 4203 participants for SARS-CoV-2 antibodies via three antibody tests. Latent class analysis yielded 8.6% standardized county-wide seroprevalence, a factor of underreported infections of 5.0, and 2.5% overall IFR. Seroprevalence was two-fold higher among medical workers and one third among current smokers with similar proportions of registered infections. While seroprevalence did not show an age-trend, the factor of underreported infections was 12.2 in the young versus 1.7 for ≥85-year-old. Age-specific IFRs were <0.5% below 60 years of age, 1.0% for age 60–69, and 13.2% for age 70+. Senior care homes accounted for 45% of COVID-19-related deaths, reflected by an IFR of 7.5% among individuals aged 70+ and an overall IFR of 1.4% when excluding senior care home residents from our computation. Our data underscore senior care home infections as key determinant of IFR additionally to age, insufficient targeted testing in the young, and the need for further investigations on behavioral or molecular causes of the fewer infections among current smokers.
Ralf Wagner; David Peterhoff; Stephanie Beileke; Felix Günther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus Stark; André Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette Pfahlberg; Iris Heid; Olaf Gefeller; Klaus Überla. Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020. Viruses 2021, 13, 1118 .
AMA StyleRalf Wagner, David Peterhoff, Stephanie Beileke, Felix Günther, Melanie Berr, Sebastian Einhauser, Anja Schütz, Hans Niller, Philipp Steininger, Antje Knöll, Matthias Tenbusch, Clara Maier, Klaus Korn, Klaus Stark, André Gessner, Ralph Burkhardt, Michael Kabesch, Holger Schedl, Helmut Küchenhoff, Annette Pfahlberg, Iris Heid, Olaf Gefeller, Klaus Überla. Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020. Viruses. 2021; 13 (6):1118.
Chicago/Turabian StyleRalf Wagner; David Peterhoff; Stephanie Beileke; Felix Günther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus Stark; André Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette Pfahlberg; Iris Heid; Olaf Gefeller; Klaus Überla. 2021. "Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020." Viruses 13, no. 6: 1118.
The response of the adaptive immune system is augmented by multimeric presentation of a specific antigen, resembling viral particles. Several vaccines have been designed based on natural or designed protein scaffolds, which exhibited a potent adaptive immune response to antigens; however, antibodies are also generated against the scaffold, which may impair subsequent vaccination. In order to compare polypeptide scaffolds of different size and oligomerization state with respect to their efficiency, including anti-scaffold immunity, we compared several strategies of presentation of the RBD domain of the SARS-CoV-2 spike protein, an antigen aiming to generate neutralizing antibodies. A comparison of several genetic fusions of RBD to different nanoscaffolding domains (foldon, ferritin, lumazine synthase, and β-annulus peptide) delivered as DNA plasmids demonstrated a strongly augmented immune response, with high titers of neutralizing antibodies and a robust T-cell response in mice. Antibody titers and virus neutralization were most potently enhanced by fusion to the small β-annulus peptide scaffold, which itself triggered a minimal response in contrast to larger scaffolds. The β-annulus fused RBD protein increased residence in lymph nodes and triggered the most potent viral neutralization in immunization by a recombinant protein. Results of the study support the use of a nanoscaffolding platform using the β-annulus peptide for vaccine design.
Duško Lainšček; Tina Fink; Vida Forstnerič; Iva Hafner-Bratkovič; Sara Orehek; Žiga Strmšek; Mateja Manček-Keber; Peter Pečan; Hana Esih; Špela Malenšek; Jana Aupič; Petra Dekleva; Tjaša Plaper; Sara Vidmar; Lucija Kadunc; Mojca Benčina; Neža Omersa; Gregor Anderluh; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Valter Bergant; Alexander Herrmann; Andreas Pichlmair; Roman Jerala. A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response. Vaccines 2021, 9, 431 .
AMA StyleDuško Lainšček, Tina Fink, Vida Forstnerič, Iva Hafner-Bratkovič, Sara Orehek, Žiga Strmšek, Mateja Manček-Keber, Peter Pečan, Hana Esih, Špela Malenšek, Jana Aupič, Petra Dekleva, Tjaša Plaper, Sara Vidmar, Lucija Kadunc, Mojca Benčina, Neža Omersa, Gregor Anderluh, Florence Pojer, Kelvin Lau, David Hacker, Bruno Correia, David Peterhoff, Ralf Wagner, Valter Bergant, Alexander Herrmann, Andreas Pichlmair, Roman Jerala. A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response. Vaccines. 2021; 9 (5):431.
Chicago/Turabian StyleDuško Lainšček; Tina Fink; Vida Forstnerič; Iva Hafner-Bratkovič; Sara Orehek; Žiga Strmšek; Mateja Manček-Keber; Peter Pečan; Hana Esih; Špela Malenšek; Jana Aupič; Petra Dekleva; Tjaša Plaper; Sara Vidmar; Lucija Kadunc; Mojca Benčina; Neža Omersa; Gregor Anderluh; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Valter Bergant; Alexander Herrmann; Andreas Pichlmair; Roman Jerala. 2021. "A Nanoscaffolded Spike-RBD Vaccine Provides Protection against SARS-CoV-2 with Minimal Anti-Scaffold Response." Vaccines 9, no. 5: 431.
Serological testing is crucial in detection of previous infection and in monitoring convalescent and vaccine-induced immunity. During the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) pandemic, numerous assay platforms have been developed and marketed for clinical use. Several studies recently compared clinical performance of a limited number of serological tests, but broad comparative evaluation is currently missing. Within this study, a panel of 161 sera from SARS-CoV-2 infected, seasonal CoV-infected and SARS-CoV-2 naïve subjects was enrolled to evaluate 16 ELISA/ECLIA-based and 16 LFA-based tests. Specificities of all ELISA/ECLIA-based assays were acceptable and generally in agreement with the providers’ specifications, but sensitivities were lower as specified. Results of the LFAs were less accurate as compared to the ELISAs, albeit with some exceptions. We found a sporadic unequal immune response for different antigens and thus recommend the use of a nucleocapsid protein (N)- and spike protein (S)-based test combination when maximal sensitivity is necessary. Finally, the quality of the immune response in terms of neutralization should be tested using S-based IgG tests.
Maren Werner; Philip Pervan; Vivian Glück; Florian Zeman; Michael Koller; Ralph Burkhardt; Thomas Glück; Jürgen Wenzel; Barbara Schmidt; André Gessner; David Peterhoff. Evaluation of a Broad Panel of SARS-CoV-2 Serological Tests for Diagnostic Use. Journal of Clinical Medicine 2021, 10, 1580 .
AMA StyleMaren Werner, Philip Pervan, Vivian Glück, Florian Zeman, Michael Koller, Ralph Burkhardt, Thomas Glück, Jürgen Wenzel, Barbara Schmidt, André Gessner, David Peterhoff. Evaluation of a Broad Panel of SARS-CoV-2 Serological Tests for Diagnostic Use. Journal of Clinical Medicine. 2021; 10 (8):1580.
Chicago/Turabian StyleMaren Werner; Philip Pervan; Vivian Glück; Florian Zeman; Michael Koller; Ralph Burkhardt; Thomas Glück; Jürgen Wenzel; Barbara Schmidt; André Gessner; David Peterhoff. 2021. "Evaluation of a Broad Panel of SARS-CoV-2 Serological Tests for Diagnostic Use." Journal of Clinical Medicine 10, no. 8: 1580.
SARS-CoV-2 infection fatality ratios (IFR) remain controversially discussed with implications for political measures, but the number of registered infections depends on testing strategies and deduced case fatality ratios (CFR) are poor proxies for IFR. The German county of Tirschenreuth suffered a severe SARS-CoV-2 outbreak in spring 2020 with particularly high CFR. To estimate seroprevalence, dark figure, and IFR for the Tirschenreuth population aged ≥14 years in June/July 2020 with misclassification error control, we conducted a population-based study, including home visits for elderly, and analyzed 4203 participants for SARS-CoV-2 antibodies via three antibody tests (64% of our random sample). Latent class analysis yielded 8.6% standardized county-wide seroprevalence, dark figure factor 5.0, and 2.5% overall IFR. Seroprevalence was two-fold higher among medical workers and one third among current smokers with similar proportions of registered infections. While seroprevalence did not show an age-trend, the dark figure was 12.2 in the young versus 1.7 for ≥85-year-old. Age-specific IFRs were <0.5% below 60 years of age, 1.0% for age 60-69, 13.2% for age 70+, confirming a previously reported age-model for IFR. Senior care homes accounted for 45% of COVID-19-related deaths, reflected by an IFR of 7.5% among individuals aged 70+ and an overall IFR of 1.4% when excluding senior care home residents from our computation. Our data underscore senior care home infections as key determinant of IFR additionally to age, insufficient targeted testing in the young, and the need for further investigations on behavioral or molecular causes of the fewer infections among current smokers.
Ralf Wagner; David Peterhoff; Stephanie Beileke; Felix Guenther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Helmut Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus J. Stark; Andre Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette B. Pfahlberg; Iris M. Heid; Olaf Gefeller; Klaus Überla. Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020. 2021, 1 .
AMA StyleRalf Wagner, David Peterhoff, Stephanie Beileke, Felix Guenther, Melanie Berr, Sebastian Einhauser, Anja Schütz, Hans Helmut Niller, Philipp Steininger, Antje Knöll, Matthias Tenbusch, Clara Maier, Klaus Korn, Klaus J. Stark, Andre Gessner, Ralph Burkhardt, Michael Kabesch, Holger Schedl, Helmut Küchenhoff, Annette B. Pfahlberg, Iris M. Heid, Olaf Gefeller, Klaus Überla. Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020. . 2021; ():1.
Chicago/Turabian StyleRalf Wagner; David Peterhoff; Stephanie Beileke; Felix Guenther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Helmut Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus J. Stark; Andre Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette B. Pfahlberg; Iris M. Heid; Olaf Gefeller; Klaus Überla. 2021. "Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020." , no. : 1.
Objective To follow serological immune responses of front-line healthcare workers after PCR-confirmed COVID-19 for a mean of 30 weeks, describe the time-course of SARS-CoV-2 spike protein-specific IgG, IgA and IgM levels and to identify associations of the immune response with symptoms, demographic parameters and severity of disease. Methods Anti-SARS-CoV-2 S protein-specific IgG, IgA and IgM antibodies were measured at three time points during the 30-week follow-up. COVID-19-specific symptoms were assessed with standardized questionnaires. Results 95% of the participants mounted an IgG response with only modest decline after week 12. IgG-type antibodies were still detectable in almost 90% of the subjects at 30 weeks. IgA and IgM responses were less robust and antibody titers decreased more rapidly. At 30 weeks, only 25% still had detectable IgA-type and none had IgM-type antibodies. Higher age and higher disease severity were independently associated with higher IgG antibody levels, albeit with wide variations. Conclusion Serological immune responses after COVID-19 show considerable inter-individual variability, but show an association with increasing age and higher severity of disease. IgG-type anti-SARS-CoV-2 antibodies remain positive in 90% of the individuals 30 weeks after onset of symptoms.
Vivian Glück; Sonja Grobecker; Leonid Tydykov; Bernd Salzberger; Thomas Glück; Tanja Weidlich; Manuela Bertok; Christine Gottwald; Jürgen J. Wenzel; André Gessner; Barbara Schmidt; David Peterhoff. SARS-CoV-2-directed antibodies persist for more than six months in a cohort with mild to moderate COVID-19. Infection 2021, 49, 739 -746.
AMA StyleVivian Glück, Sonja Grobecker, Leonid Tydykov, Bernd Salzberger, Thomas Glück, Tanja Weidlich, Manuela Bertok, Christine Gottwald, Jürgen J. Wenzel, André Gessner, Barbara Schmidt, David Peterhoff. SARS-CoV-2-directed antibodies persist for more than six months in a cohort with mild to moderate COVID-19. Infection. 2021; 49 (4):739-746.
Chicago/Turabian StyleVivian Glück; Sonja Grobecker; Leonid Tydykov; Bernd Salzberger; Thomas Glück; Tanja Weidlich; Manuela Bertok; Christine Gottwald; Jürgen J. Wenzel; André Gessner; Barbara Schmidt; David Peterhoff. 2021. "SARS-CoV-2-directed antibodies persist for more than six months in a cohort with mild to moderate COVID-19." Infection 49, no. 4: 739-746.
Effective and safe vaccines against SARS-CoV-2 are highly desirable to prevent casualties and societal cost caused by Covid-19 pandemic. The receptor binding domain (RBD) of the surface-exposed spike protein of SARS-CoV-2 represents a suitable target for the induction of neutralizing antibodies upon vaccination. Small protein antigens typically induce weak immune response while particles measuring tens of nanometers are efficiently presented to B cell follicles and subsequently to follicular germinal center B cells in draining lymph nodes, where B cell proliferation and affinity maturation occurs. Here we prepared and analyzed the response to several DNA vaccines based on genetic fusions of RBD to four different scaffolding domains, namely to the foldon peptide, ferritin, lumazine synthase and β-annulus peptide, presenting from 6 to 60 copies of the RBD on each particle. Scaffolding strongly augmented the immune response with production of neutralizing antibodies and T cell response including cytotoxic lymphocytes in mice upon immunization with DNA plasmids. The most potent response was observed for the 24-residue β-annulus peptide scaffold that forms large soluble assemblies, that has the advantage of low immunogenicity in comparison to larger scaffolds. Our results support the advancement of this vaccine platform towards clinical trials.
Dusko Lainscek; Tina Fink; Vida Forstneric; Iva Hafner-Bratkovic; Sara Orehek; Ziga Strmsek; Mateja Mancek Keber; Peter Pecan; Hana Esih; Spela Malensek; Jana Aupic; Petra Dekleva; Tjasa Plaper; Sara Vidmar; Lucija Kadunc; Mojca Bencina; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Roman Jerala. Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein. 2020, 1 .
AMA StyleDusko Lainscek, Tina Fink, Vida Forstneric, Iva Hafner-Bratkovic, Sara Orehek, Ziga Strmsek, Mateja Mancek Keber, Peter Pecan, Hana Esih, Spela Malensek, Jana Aupic, Petra Dekleva, Tjasa Plaper, Sara Vidmar, Lucija Kadunc, Mojca Bencina, Florence Pojer, Kelvin Lau, David Hacker, Bruno Correia, David Peterhoff, Ralf Wagner, Roman Jerala. Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein. . 2020; ():1.
Chicago/Turabian StyleDusko Lainscek; Tina Fink; Vida Forstneric; Iva Hafner-Bratkovic; Sara Orehek; Ziga Strmsek; Mateja Mancek Keber; Peter Pecan; Hana Esih; Spela Malensek; Jana Aupic; Petra Dekleva; Tjasa Plaper; Sara Vidmar; Lucija Kadunc; Mojca Bencina; Florence Pojer; Kelvin Lau; David Hacker; Bruno Correia; David Peterhoff; Ralf Wagner; Roman Jerala. 2020. "Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein." , no. : 1.
Objective The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels. Methods In our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated. Results The RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R 2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%; N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection; N = 53) in serum. Conclusions With the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination.
David Peterhoff; Vivian Glück; Matthias Vogel; Philipp Schuster; Anja Schütz; Philip Neubert; Veruschka Albert; Stefanie Frisch; Mara Kiessling; Philip Pervan; Maren Werner; Nicole Ritter; Leon Babl; Maria Deichner; Frank Hanses; Matthias Lubnow; Thomas Müller; Dirk Lunz; Florian Hitzenbichler; Franz Audebert; Viola Hähnel; Robert Offner; Martina Müller; Stephan Schmid; Ralph Burkhardt; Thomas Glück; Michael Koller; Hans Helmut Niller; Bernhard Graf; Bernd Salzberger; Jürgen Wenzel; Jonathan Jantsch; André Gessner; Barbara Schmidt; Ralf Wagner. A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization. Infection 2020, 49, 75 -82.
AMA StyleDavid Peterhoff, Vivian Glück, Matthias Vogel, Philipp Schuster, Anja Schütz, Philip Neubert, Veruschka Albert, Stefanie Frisch, Mara Kiessling, Philip Pervan, Maren Werner, Nicole Ritter, Leon Babl, Maria Deichner, Frank Hanses, Matthias Lubnow, Thomas Müller, Dirk Lunz, Florian Hitzenbichler, Franz Audebert, Viola Hähnel, Robert Offner, Martina Müller, Stephan Schmid, Ralph Burkhardt, Thomas Glück, Michael Koller, Hans Helmut Niller, Bernhard Graf, Bernd Salzberger, Jürgen Wenzel, Jonathan Jantsch, André Gessner, Barbara Schmidt, Ralf Wagner. A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization. Infection. 2020; 49 (1):75-82.
Chicago/Turabian StyleDavid Peterhoff; Vivian Glück; Matthias Vogel; Philipp Schuster; Anja Schütz; Philip Neubert; Veruschka Albert; Stefanie Frisch; Mara Kiessling; Philip Pervan; Maren Werner; Nicole Ritter; Leon Babl; Maria Deichner; Frank Hanses; Matthias Lubnow; Thomas Müller; Dirk Lunz; Florian Hitzenbichler; Franz Audebert; Viola Hähnel; Robert Offner; Martina Müller; Stephan Schmid; Ralph Burkhardt; Thomas Glück; Michael Koller; Hans Helmut Niller; Bernhard Graf; Bernd Salzberger; Jürgen Wenzel; Jonathan Jantsch; André Gessner; Barbara Schmidt; Ralf Wagner. 2020. "A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization." Infection 49, no. 1: 75-82.
The rise of antibiotic resistance demands the acceleration of molecular diversification strategies to inspire new chemical entities for antibiotic medicines. We report here on the large-scale engineering of ribosomally synthesized and post-translationally modified antimicrobial peptides carrying the ring-forming amino acid lanthionine. New-to-nature variants featuring distinct properties were obtained by combinatorial shuffling of peptide modules derived from 12 natural antimicrobial lanthipeptides and processing by a promiscuous post-translational modification machinery. For experimental characterization, we developed the nanoFleming, a miniaturized and parallelized high-throughput inhibition assay. On the basis of a hit set of >100 molecules, we identified variants with improved activity against pathogenic bacteria and shifted activity profiles, and extrapolated design guidelines that will simplify the identification of peptide-based anti-infectives in the future.
Steven Schmitt; Manuel Montalbán-López; David Peterhoff; Jingjing Deng; Ralf Wagner; Martin Held; Oscar P. Kuipers; Sven Panke. Analysis of modular bioengineered antimicrobial lanthipeptides at nanoliter scale. Nature Chemical Biology 2019, 15, 437 -443.
AMA StyleSteven Schmitt, Manuel Montalbán-López, David Peterhoff, Jingjing Deng, Ralf Wagner, Martin Held, Oscar P. Kuipers, Sven Panke. Analysis of modular bioengineered antimicrobial lanthipeptides at nanoliter scale. Nature Chemical Biology. 2019; 15 (5):437-443.
Chicago/Turabian StyleSteven Schmitt; Manuel Montalbán-López; David Peterhoff; Jingjing Deng; Ralf Wagner; Martin Held; Oscar P. Kuipers; Sven Panke. 2019. "Analysis of modular bioengineered antimicrobial lanthipeptides at nanoliter scale." Nature Chemical Biology 15, no. 5: 437-443.