A peptide-based assay discriminates individual antibody response to SARS-CoV-2

Polvere, Immacolata; Voccola, Serena; Cardinale, Gaetano; Fumi, Maurizio; Aquila, F; Parrella, Alfredina; Madera, Jessica Raffaella; Stilo, Romania; Vito, Pasquale; Zotti, Tiziana

doi:10.1016/j.gendis.2021.01.008

Cited by 12 publications

(18 citation statements)

References 17 publications

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“…Out of 63, only five samples resulted positive, four of which were confirmed by an IVD-validated kit. This definitely low number of SARS-CoV-2-positive individuals identified in the community of the University of Sannio at the beginning of the academic year is consistent with previous seroprevalence studies indicating that early restrictions were successful in limiting COVID-19 diffusion in the district of Benevento [ 24 , 25 ].…”

Section: Discussionsupporting

confidence: 89%

Sample-Pooling Strategy for SARS-CoV-2 Detection among Students and Staff of the University of Sannio

et al. 2021

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Since the beginning of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic, it has been clear that testing large groups of the population was the key to stem infection and prevent the effects of the coronavirus disease of 2019, mostly among sensitive patients. On the other hand, time and cost-sustainability of virus detection by molecular analysis such as reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) may be a major issue if testing is extended to large communities, mainly asymptomatic large communities. In this context, sample-pooling and test grouping could offer an effective solution. Here we report the screening on 1195 oral-nasopharyngeal swabs collected from students and staff of the Università degli Studi del Sannio (University of Sannio, Benevento, Campania, Italy) and analyzed by an in-house developed multiplex RT-qPCR for SARS-CoV-2 detection through a simple monodimensional sample pooling strategy. Overall, 400 distinct pools were generated and, within 24 h after swab collection, five positive samples were identified. Out of them, four were confirmed by using a commercially available kit suitable for in vitro diagnostic use (IVD). High accuracy, sensitivity and specificity were also determined by comparing our results with a reference IVD assay for all deconvoluted samples. Overall, we conducted 463 analyses instead of 1195, reducing testing resources by more than 60% without lengthening diagnosis time and without significant losses in sensitivity, suggesting that our strategy was successful in recognizing positive cases in a community of asymptomatic individuals with minor requirements of reagents and time when compared to normal testing procedures.

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Section: Discussionsupporting

confidence: 89%

Sample-Pooling Strategy for SARS-CoV-2 Detection among Students and Staff of the University of Sannio

et al. 2021

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“…The N-terminus (aa 1 to~14) is proposed to be exposed to the exterior of the virus, while the long C-terminal portion of M (following aa~85-99), representing about half of the amino acid residues, is proposed to be inside the viral particle (7)(8)(9). Epitopes residing within the N-terminal end of SARS-CoV-2 have also been observed by others (26,33,35). The N-terminus of the M protein contains the only N-glycosylation site of the protein, a highly conserved residue among coronaviruses.…”

Section: Discussionmentioning

confidence: 74%

“…From our panel of 21 peptides only two peptides, stemming from the very N- and C-terminal ends of the M protein, peptides M01 and M21, were specifically recognized by antibodies of COVID-19 patients, while the other peptides did not provoke specific antibody reactivity as compared to historical plasma samples collected before the pandemic. In contrast to other peptide-based B-cell epitope mapping analyses of the coronavirus M protein ( 19 , 32 , 33 ), we and others employed the biotin–streptavidin system for peptide coating ( 21 , 26 , 32 ). The proposed topology of the M protein may however elucidate the observed antibody reactivity.…”

Section: Discussionmentioning

confidence: 99%

Antibody Response to SARS-CoV-2 Membrane Protein in Patients of the Acute and Convalescent Phase of COVID-19

et al. 2021

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Understanding the course of the antibody response directed to individual epitopes of SARS-CoV-2 proteins is crucial for serological assays and establishment of vaccines. Twenty-one synthetic peptides were synthesized that have ten amino acids overlap and cover the complete membrane (M) protein. Plasma samples from 32 patients having acute disease and 30 patients from the convalescent phase were studied. Only peptide M01 (aa 1–20) and to a lesser extent peptide M21 (aa 201–222) showed specific reactivity as compared to historical control plasma samples. Peptide M01 was recognized by IgM- (71.9%) and IgG-specific antibodies (43.8%) during the acute phase as early as day 8 PIO. In a longitudinal analysis, a higher reactivity was observed for the IgM response directed to peptide M01 following day 20 PIO as compared to earlier time points of the acute phase. In the convalescent phase, antibody reactivity to the two M-specific peptides was significantly lower (<30% seropositivity). A fusion protein encoding major parts of RBD also showed higher rates of recognition during acute (50.0%) and lower rates in the convalescent phase (23.3%). Taken together, our results suggest that during the acute phase of COVID-19 antibodies are raised to two linear epitopes of the SARS-CoV-2 M protein, located at the very N- and C-termini, showing almost similar levels of reactivity as immunodominant linear epitopes derived from the spike and nucleocapsid protein. Anti-M is also present in the convalescent phase of COVID-19 patients, however at lower levels, with the N-terminus of the M protein as a preferred target.

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“…Antibody mapping has the potential to further define relevant antigens [136][137][138]. Microarrays offer a more complex mechanism to define immunoreactive epitopes even when honed down to a few larger but common antigens such as S and N proteins [42,139]. Neutralizing antibody levels may also correlate well with particular adaptive T cell responses [111].…”

Section: Correlations With Immunity or Protectionmentioning

confidence: 99%

Passive Immunity Should and Will Work for COVID-19 for Some Patients

Cimolai¹

2021

CHI

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In the absence of effective antiviral chemotherapy and still in the context of emerging vaccines for severe acute respiratory syndrome-CoV-2 infections, passive immunotherapy remains a key treatment and possible prevention strategy. What might initially be conceived as a simplified donor-recipient process, the intricacies of donor plasma, IV immunoglobulins, and monoclonal antibody modality applications are becoming more apparent. Key targets of such treatment have largely focused on virus neutralization and the specific viral components of the attachment Spike protein and its constituents (e.g., receptor binding domain, N-terminal domain). The cumulative laboratory and clinical experience suggests that beneficial protective and treatment outcomes are possible. Both a dose-and a time-dependency emerge. Lesser understood are the concepts of bioavailability and distribution. Apart from direct antigen binding from protective immunoglobulins, antibody effector functions have potential roles in outcome. In attempting to mimic the natural but variable response to infection or vaccination, a strong functional polyclonal approach attracts the potential benefits of attacking antigen diversity, high antibody avidity, antibody persistence, and protection against escape viral mutation. The availability and ease of administration for any passive immunotherapy product must be considered in the current climate of need. There is never a perfect product, but yet there is considerable room for improving patient outcomes. Given the variability of human genetics, immunity, and disease, and given the nuances of the virus and its potential for change, passive immunotherapy can be developed that will be effective for some but not all patients. An understanding of such patient variability and limitations is just as important as the understanding of the direct interactions between immunotherapy and virus.

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A peptide-based assay discriminates individual antibody response to SARS-CoV-2

Cited by 12 publications

References 17 publications

Sample-Pooling Strategy for SARS-CoV-2 Detection among Students and Staff of the University of Sannio

Sample-Pooling Strategy for SARS-CoV-2 Detection among Students and Staff of the University of Sannio

Antibody Response to SARS-CoV-2 Membrane Protein in Patients of the Acute and Convalescent Phase of COVID-19

Passive Immunity Should and Will Work for COVID-19 for Some Patients

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