SARS-CoV-2 Antibody Binding and Neutralization in Dried Blood Spot Eluates and Paired Plasma

Itell, Hannah L.; Weight, Haidyn; Fish, Carolyn S.; Logue, Jennifer; Franko, Nicholas; Wolf, Caitlin R; McCulloch, Denise J.; Galloway, Jared; Matsen, F. A.; Overbaugh, Julie

doi:10.1128/spectrum.01298-21

Cited by 24 publications

(28 citation statements)

References 44 publications

(72 reference statements)

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“…Overall, the study results show a correlation of r > 0.96 for DBS and serum for both SARS-CoV-2 antibodies, S1 and NCP. Our results are in accordance with other work, presenting similarly high correlation for both materials regarding SARS-CoV-2 antibody analysis [ 13 , [16] , [17] , [18] ]. Suitability of DBS for analysis of SARS-CoV-2 antibodies was also shown using assays from other suppliers [ 19 , 20 ], as well as in-house methods or manual testing [ 9 , 21 , 22 ].…”

Section: Discussionsupporting

confidence: 93%

Dried blood spot eluates are suitable for testing of SARS-CoV-2 IgG antibodies targeting Spike protein 1 and Nucleocapsid protein

Guttmann

Bunte

Eschrig

et al. 2023

Biochemistry and Biophysics Reports

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Section: Discussionsupporting

confidence: 93%

Dried blood spot eluates are suitable for testing of SARS-CoV-2 IgG antibodies targeting Spike protein 1 and Nucleocapsid protein

Guttmann

Bunte

Eschrig

et al. 2023

Biochemistry and Biophysics Reports

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“…Direct ELISA : The binding affinities of the mAbs to recombinant stabilized spike trimers were assessed by direct ELISA as previously described (54, 78). The recombinant spike trimers used in these assays were as follows: Wuhan-Hu-1 (Sino Biologics, cat.…”

Section: Methodsmentioning

confidence: 99%

“…Neutralization of the pseudoviruses by the plasma and mAbs was performed as previously described (54, 55). HEK293T-ACE2 cells were seeded at ∼4000 cells/well in DMEM (supplemented with 10% FBS and penicillin/streptomycin/fungizone) in a black-walled 384 well plate.…”

Section: Methodsmentioning

confidence: 99%

Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection

Guenthoer

Lilly

Starr

et al. 2022

Preprint

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The antiviral benefit of antibodies can be compromised by viral escape especially for rapidly evolving viruses. Therefore, durable, effective antibodies must be both broad and potent to counter newly emerging, diverse strains. Discovery of such antibodies is critically important for SARS-CoV-2 as the global emergence of new variants of concern (VOC) has compromised the efficacy of therapeutic antibodies and vaccines. We describe a collection of broad and potent neutralizing monoclonal antibodies (mAbs) isolated from an individual who experienced a breakthrough infection with the Delta VOC. Four mAbs potently neutralize the Wuhan-Hu-1 vaccine strain, the Delta VOC, and also retain potency against the Omicron VOCs, including recently circulating BA.4/BA.5, in both pseudovirus-based and live virus assays, and one also potently neutralizes SARS-CoV-1. The potency of these mAbs was greater against Omicron VOCs than all but one of the mAbs that had been approved for therapeutic applications. The mAbs target distinct epitopes on the spike glycoprotein, three in the receptor binding domain (RBD) and one in an invariant region downstream of the RBD in subdomain 1 (SD1). The escape pathways we defined at single amino acid resolution with deep mutational scanning show they target conserved, functionally constrained regions of the glycoprotein, suggesting escape could incur a fitness cost. Overall, these mAbs are novel in their breadth across VOCs, their epitope specificity, and include a highly potent mAb targeting a rare epitope outside of the RBD in SD1.

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“…We hypothesized that higher antibody binding levels against full-length Spike, RBD, NTD and FP in infants might correlate with elevated functional antibody activity, including neutralization and/or ADCC. We therefore tested plasma in a Spike-pseudotyped lentiviral neutralization assay (Crawford et al, 2020; Itell et al, 2021). Mothers living with HIV and HIV-exposed infants were excluded from the neutralization analysis because of the presence of ART in plasma samples, which would inhibit infection in the assay.…”

Section: Resultsmentioning

confidence: 99%

“…384-well format SARS-CoV-2 Spike neutralization assays SARS-CoV-2 Spike-pseudotyped lentiviral neutralization assays were conducted in a 384-well plate format as described previously (Itell et al, 2021). Briefly, black-walled, clear bottom, poly-L-lysinecoated 384-well plates (Thermo Scientific #142761) were seeded with 3.75 x 10 3 HEK293T-ACE2 cells (BEI Resources, NR-52511) per well in 30 µL of complete DMEM.…”

Section: Sars-cov-2 Spike-pseudotyped Lentivirus Productionmentioning

confidence: 99%

Elevated binding and functional antibody responses to SARS-CoV-2 in infants versus mothers

Stoddard

Sung

Yaffe

et al. 2023

Preprint

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Infant antibody responses to viral infection can differ from those in adults. However, data on the specificity and function of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in infants, and direct comparisons between infants and adults are limited. We characterized antibody binding and functionality in convalescent plasma from postpartum women and their infants infected with SARS-CoV-2 from a vaccine-naive prospective cohort in Nairobi, Kenya. Antibody titers against SARS-CoV-2 Spike, receptor binding domain and N-terminal domain, and Spike-expressing cell-surface staining levels were significantly higher in infants than in mothers. Plasma antibodies from mothers and infants bound to similar regions of the Spike S2 subunit, including the fusion peptide (FP) and stem helix-heptad repeat 2. However, infants displayed higher antibody levels and more consistent antibody escape pathways in the FP region compared to mothers. Finally, infants had significantly higher levels of antibody-dependent cellular cytotoxicity (ADCC), though, surprisingly, neutralization titers between infants and mothers were similar. These results suggest infants develop distinct SARS-CoV-2 binding and functional antibody repertoires and reveal age-related differences in humoral immunity to SARS-CoV-2 infection that could be relevant to protection and COVID-19 disease outcomes.

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SARS-CoV-2 Antibody Binding and Neutralization in Dried Blood Spot Eluates and Paired Plasma

Abstract: Plasma and sera isolated from venous blood represent conventional sample types used for the evaluation of SARS-CoV-2 antibody responses after infection or vaccination. However, collection of these samples is invasive and requires trained personnel and equipment for immediate processing.

Cited by 24 publications

References 44 publications

Dried blood spot eluates are suitable for testing of SARS-CoV-2 IgG antibodies targeting Spike protein 1 and Nucleocapsid protein

Dried blood spot eluates are suitable for testing of SARS-CoV-2 IgG antibodies targeting Spike protein 1 and Nucleocapsid protein

Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection

Elevated binding and functional antibody responses to SARS-CoV-2 in infants versus mothers

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