Journal: Virologica Sinica

Article Title: Omicron variants breakthrough infection elicited higher specific memory immunity than third dose booster in healthy vaccinees.

doi: 10.1016/j.virs.2022.12.008

Figure Lengend Snippet: Fig. 1. Comparative analysis of anti-ancestral-spike-trimer-IgG. Serum were collected from vaccinated healthy volunteers (HVs) before third booster, at median of 33 days post inactivated vaccine (InV) booster and 30.5 days post Ad5-nCoV booster, and Omicron variants breakthrough infected patients (OBIPs) at median of 32 days post infection. A Anti-spike-trimer-IgG titer in InV boosted HVs, Ad5-nCoV boosted HVs, and OBIPs were determined by ELISA. B–G Comparative analysis of SARS- CoV-2 ancestral strain spike-trimer specific IgG subclass. Anti-spike-trimer-IgG1 (B), anti-Spike-trimer-IgG2 (C), anti-Spike-trimer-IgG3 (D), and anti-spike-trimer-IgG4 (E) titer in InV boosted HVs, Ad5-nCoV boosted HVs, and OBIPs were determined by ELISA. F The ratio of spike-trimer specific IgG1/IgG4 in InV boosted HVs, Ad5- nCoV boosted HVs, and OBIPs. G The ratio of spike-trimer specific IgG1þIgG3/IgG4þIgG2 in InV boosted HVs, Ad5-nCoV boosted HVs, and OBIPs. Data presented as geometric mean titers (GMT) and 95% confidence interval. Dotted line: the detection limits. Statistical analyses were performed by Mann-Whitney U test.

Article Snippet: After blocking, 100 μL 3-fold diluted plasma (initially dilute for IgG, IgG1, IgG2 and IgG3:1:20, IgG4:1:10) was added in each well, and incubated at 37 C for 1 h. After thorough washing, 100 μL HRP labeled secondary antibodies against human IgG (Cat#: 2010-05, 1:10,000, Southern Biotech, USA), IgG1 (Cat#: 9054-05, 1:4000, Southern Biotech, USA), IgG2 (Cat#: 9060- 05, 1:2000, Southern Biotech, USA), IgG3 (Cat#: 9210-05, 1:4000, Southern biotech, USA), and IgG4 (Cat#: 9200-05, 1:4000, Southern Biotech, USA) was added in each well.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Virologica Sinica

Article Title: Omicron variants breakthrough infection elicited higher specific memory immunity than third dose booster in healthy vaccinees.

doi: 10.1016/j.virs.2022.12.008

Figure Lengend Snippet: Fig. 2. Comparative analysis of anti-RBD antibody against SARS-CoV-2. Serum were collected vaccinated healthy volunteers (HVs) before third booster, at median of 33 days post InV booster and 30.5 days post Ad5-nCoV booster, and OBIPs at median of 32 days post infection. Anti-ancestral-receptor binding domain (RBD)-IgG (A), anti-Delta-RBD-IgG (B), and anti-Omicron-RBD-IgG (C) titer in pre-boosted HVs, InV boosted HVs, Ad5-nCoV boosted HVs, and OBIPs were determined by ELISA. D Paired analysis of anti-ancestral-RBD-, anti-Delta-RBD-, and anti-Omicron-RBD-IgG titer in pre-boosted HVs, InV boosted HVs, Ad5-nCoV boosted HVs, and OBIPs. Data presented as geometric mean titers (GMT) and 95% confidence interval. Dotted line: the detection limits. Statistical analyses were performed by Mann-Whitney U test.

Article Snippet: After blocking, 100 μL 3-fold diluted plasma (initially dilute for IgG, IgG1, IgG2 and IgG3:1:20, IgG4:1:10) was added in each well, and incubated at 37 C for 1 h. After thorough washing, 100 μL HRP labeled secondary antibodies against human IgG (Cat#: 2010-05, 1:10,000, Southern Biotech, USA), IgG1 (Cat#: 9054-05, 1:4000, Southern Biotech, USA), IgG2 (Cat#: 9060- 05, 1:2000, Southern Biotech, USA), IgG3 (Cat#: 9210-05, 1:4000, Southern biotech, USA), and IgG4 (Cat#: 9200-05, 1:4000, Southern Biotech, USA) was added in each well.

Techniques: Infection, Binding Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Costimulatory receptors in a teleost fish: typical CD28, elusive CTLA4.

doi: 10.4049/jimmunol.176.7.4191

Figure Lengend Snippet: FIGURE 8. Combined triggering of CD3 and huCD28/rbtCD28 in- duces IL-2 production. Jurkat CH7C17 transfectants expressing huCD28 wild-type (A14), huCD28/rbtCD28 receptor (2F5), huCD28/ rbtCTLA4 receptor (26H6) were incubated with ( ) or without (f) anti-human CD28 Ab for 30 min at 4°C. After washing, cells were deposited on anti-CD3-coated plates, and goat anti-mouse IgG1 Ab was added to cross-link human CD28 or chimeric receptors. Cells incubated only with goat anti-mouse IgG1 Ab were used as a control (). Su- pernatants were collected after 24 h at 37°C, and the IL-2 titer was determined by ELISA. IL-2 concentration is indicated in pg/10 ml. Asterisks indicate that the production of IL-2 by cells incubated using anti-human CD3-coated plates in the presence of anti-human CD28 Ab was significantly different from the production of IL-2 by cells solely plated on anti-human CD3-coated wells (two-tailed Mann-Whitney U test; , p 0.05, , p 0.01).

Article Snippet: Goat anti-mouse IgG1 Ab ( 1 chain specific; Southern Biotechnology Associates) was added (5 g/ ml) to cross-link human CD28.

Techniques: Expressing, Incubation, Control, Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test, MANN-WHITNEY

Journal: eLife

Article Title: Glycan-based shaping of the microbiota during primate evolution

doi: 10.7554/eLife.67450

Figure Lengend Snippet:

Article Snippet: Antibody , Goat Anti-Mouse Polyclonal IgM, Human ads-HRP , Southern Biotech , Cat# 1020–05, RRID: AB_2794201 , 1:4000 (ELISA).

Techniques: Probiotics, Expressing, Bacteria, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Serial Dilution, Liposomes, Control, In Vivo, Staining, Software

Journal: Scientific Reports

Article Title: In vitro complement activation via nucleocapsid and spike proteins of SARS-CoV-2 in COVID-19 patients

doi: 10.1038/s41598-025-20926-6

Figure Lengend Snippet: Comparison of the in vitro C3b and C4b complement depositions via spike (C3b(S) and C4b(S)) or nucleocapsid (C3b(N) and C4b(N)) recombinant proteins between patients with (Ab(+)) or without (Ab(-)) SARS-CoV-2-specific IgM and IgG antibodies from four different COVID-19 groups (CONV, HOSP, HOSP + O 2 and ICU). The in vitro C3b and C4b complement depositions were measured by our in-house complement deposition assays, where we used carrier free recombinant nucleocapsid or spike proteins and non-heat-inactivated patient sera. The p values for the pair-wise group comparisons on the violin plots were calculated by the Mann-Whitney tests. P values of significant differences are marked as p MW between without and with specific antibodies. Non-significant differences are marked as n.s.

Article Snippet: Mouse anti-human IgG1 Fc-HRP (HP6001) , SouthernBiotech , 9054-05.

Techniques: Comparison, In Vitro, Recombinant, MANN-WHITNEY

Journal: Scientific Reports

Article Title: In vitro complement activation via nucleocapsid and spike proteins of SARS-CoV-2 in COVID-19 patients

doi: 10.1038/s41598-025-20926-6

Figure Lengend Snippet: Comparison of the levels of SARS-CoV-2-specific IgM and IgG among four COVID-19 groups (CONV, HOSP, HOSP + O2, ICU) measured by Generic Assays CoV-2 IgM or IgG ( a ). The percentages of SARS-CoV-2-specific IgG or IgM positivity were shown also for four different COVID-19 groups. The numbers represent the mean percentages of SARS-CoV-2-specific IgM in blue or specific IgG in green circle sectors ( b ). The levels of total S- or N-specific IgG were quantified by in-house ELISA, normalized to positive and negative serum samples ( c ). The levels of S-specific and N-specific IgG1 ( d ) and IgG3 ( e ) subclasses were quantified by in-house ELISA calibrated with purified human IgG1 and IgG3. The p -values ( p KW ) were determined by Kruskal-Wallis test.

Article Snippet: Mouse anti-human IgG1 Fc-HRP (HP6001) , SouthernBiotech , 9054-05.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Purification

Journal: Scientific Reports

Article Title: In vitro complement activation via nucleocapsid and spike proteins of SARS-CoV-2 in COVID-19 patients

doi: 10.1038/s41598-025-20926-6

Figure Lengend Snippet: The in vitro C3b and C4b complement depositions via spike (C3b(S) and C4b(S)) recombinant protein are plotted by the levels of S-specific IgG1 and IgG3 in CONV individuals and in the other three hospitalized COVID-19 groups (HOSP, HOSP + O 2 and ICU) ( a ). The in vitro C3b and C4b complement depositions via nucleocapsid (C3b(N) and C4b(N)) recombinant protein are plotted by the levels of N-specific IgG1 and IgG3 in CONV individuals and in the other three hospitalized COVID-19 groups (HOSP, HOSP + O 2 and ICU) ( b ).

Article Snippet: Mouse anti-human IgG1 Fc-HRP (HP6001) , SouthernBiotech , 9054-05.

Techniques: In Vitro, Recombinant

Journal: Scientific Reports

Article Title: In vitro complement activation via nucleocapsid and spike proteins of SARS-CoV-2 in COVID-19 patients

doi: 10.1038/s41598-025-20926-6

Figure Lengend Snippet: Comparison of the levels of SARS-CoV-2-specific IgM and IgG antibodies, and in vivo complement profiles in all COVID-19 patients between low and high in vitro C3b(N) complement depositions. The p values for the pair-wise group comparisons on the violin plots were calculated by the Mann-Whitney tests. Asterisks indicate p value < 0.0143 , considered significant results after 5% FDR correction using the Benjamini-Hochberg method.

Article Snippet: Mouse anti-human IgG1 Fc-HRP (HP6001) , SouthernBiotech , 9054-05.

Techniques: Comparison, In Vivo, In Vitro, MANN-WHITNEY

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 6. In vitro antigenicity analysis of t-M2e-t NCs using ELISA. (A) Absorbance values from an indirect ELISA performed by coating 96-well plates with a fixed amount of either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a negative control peptide (NCP). Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of M2e detection IgG antibody, which was applied at different dilutions. (B) Absorbance values from a sandwich ELISA performed by coating 96-well plates with a fixed amount of purified mouse anti-M2e IgM monoclonal antibody as capture antibody. Dilutions of analyte (either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a NCP) were then added to the wells. Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of primary M2e detection IgG antibody. Cartoons of each ELISA procedure are shown below their respective graphs.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Indirect ELISA, Negative Control, Formulation, Sandwich ELISA

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 7. Immune response and survival data of mice after immunization with t-M2e-t NCs. Mice were immunized thrice, once each on day 0, 21, and 42. Blood was collected and analyzed for anti-M2e antibodies. Within 1 week of the final serum collection, mice were challenged with 3× LD50 A/California/07/2009 (H1N1) and monitored for 14 days. Two antigens were used (i) 20 μg or 5 μg t-M2e-t UCP (un-cross-linked peptide) with/without 20 μg CpG, and (ii) 20 μg or 5 μg S t-M2e-t NCs (small NCs) with/without 20 μg CpG. (A−C) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t UCP as antigen. (D−F) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t NC as antigen. (G, H) Body weight and survival after virus challenge of t-M2e-t UCP groups. (I, J) Body weight and survival after virus challenge of t-M2e-t NP groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: Virus

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against OVA and subsequent tumour inoculation of the OVA-expressing B16F10 cell lines in mice. B) Titers of anti-OVA IgG per IgG subtypes in the plasma of the mice at d18. C) Percentage of OVA-specific CD8 + T cells against the immunogenic epitope OVA 257-264 (SIINFEKL) in the blood at d18 in pre-immunised mice (vax) or naïve mice (no vax) (n ≥ 3, mean ± SD, unpaired t-test). D, E) B16F10 melanoma cells, either wild-type (WT) or genetically modified to express high ( HI ) or low ( LO ) doses of membrane-bound OVA (B16mOVA) or soluble OVA (B16-OVA), were injected intradermally in C57BL/6 mice. Tumour growth (D) and associated survival (E) of the different OVA-expressing B16 cell lines in pre-immunised mice (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d10, d14 or d26 for tumour growth, log-rank tests for survival). F) Potential anti-tumour mechanisms engaged by soluble (left) or membrane-bound (right) xenoantigens when expressed by cancer cells.

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Expressing, Clinical Proteomics, Genetically Modified, Membrane, Injection

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Immunofluorescent staining of B16F10 WT melanoma with the TRP1-targeting TA99 antibody (scale bar = 100 μm). B) B16F10 WT tumour growth upon treatment with TA99 or IgG2a isotype control (n ≥ 4, mean ± SEM, Mann-Whitney test on d11). C) Design of FabTRP-OVA fusion protein and analysis of its production by SDS-PAGE (expected size = 92,3 kDa). D) Illustration of the FabTRP-OVA fusion protein approach for targeting the xenoantigen to the cancer cell membrane. E) Representative flow cytometry plot of FabTRP-OVA, OVA and no OVA (i.e., secondary antibody only) binding to the surface of B16F10 WT cells. F) B16F10 WT tumour growth when treated with FabTRP-OVA in pre-immunised or naïve mice (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). G) B16F10 WT tumour growth when treated with FabTRP-OVA in combination with anti-PD-1 checkpoint blocade therapy (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). H) Experimental treatment timeline of xenoantigen delivery (i.e., FabTRP-OVA, OVA or PBS (vehicle only)) and anti-PD-1 checkpoint blockade therapy in B16F10 WT in mice pre-immunised against OVA (Vax) or naïve (No vax). I, J) B16F10 WT tumour growth (I) and associated mouse survival (J) upon treatment with FabTRP-OVA, OVA or PBS in combination with anti-PD-1 checkpoint blockade therapy (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Staining, Control, MANN-WHITNEY, SDS Page, Membrane, Flow Cytometry, Binding Assay

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against varicella VZVO using Varivax or a combination of gE/CpG. B) IFΝγ quantification in the supernatant of gE restimulated (+) or non-restimulated (-) splenocytes, collected from mice immunized with Varivax, gE/CpG or PBS (i.e., naïve). Ionomycin+PMA was used as a positive control. C) Titers of anti-gE total IgG and per IgG subtypes in the plasma of the pre-immunised mice at d55 (before tumour implantation). D) Experimental treatment timeline of xenoantigen delivery (i.e., Varivax, gE or PBS (vehicle only)) and anti-PD-1 checkpoint blockade treatment in B16F10 WT in mice pre-immunised with Varivax against varicella VZVO . E, F) B16F10 WT tumour growth (E) and associated mouse survival (F) upon treatment with Varivax, gE or PBS in combination with anti-PD-1 checkpoint blockade (n ≥ 7, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Positive Control, Clinical Proteomics

Journal: EBioMedicine

Article Title: Adjuvant-enhanced CD4 T Cell Responses are Critical to Durable Vaccine Immunity

doi: 10.1016/j.ebiom.2015.11.041

Figure Lengend Snippet: Adjuvants impact the durability of protection conferred by eVLP vaccination. (a) C57BL/6 mice were vaccinated IM two times with 10 μg of eVLP and challenged four (short-term) weeks or twenty-two (long-term) weeks after the vaccine boost. Data in A are pooled from 8 individual studies with 6–10 animals/group. Fisher's exact test: survival in the short-term group was significantly higher than in the long-term group (p < 0.0001). (b) C57BL/6 mice were vaccinated IM two times with 10 μg of eVLP and challenged at the indicated days after the second vaccination. n = 9 or 10/group. Cochran-Armitage test: percentage surviving declined as time to challenge increased (p = 0.0046). There was a significant difference (p = 0.03) between survival on Day 77 and Day 175. (c) Serum samples collected from animals in (B) one week prior to challenge were subjected to an ELISA for the evaluation of anti-GP IgG, IgG1, and IgG2c antibody titers. Red symbols indicate titers of animals that succumbed to challenge while black indicate titers of survivors; red symbols with black outlines indicate that one of these two animals succumbed to challenge, but animal tags were indeterminate after challenge. Median and IQR shown. (d) C57BL/6 mice were vaccinated two times (IM) with VLP, with or without the indicated adjuvants. Animals were challenged twenty-two weeks after the vaccine boost. Data in D are pooled from at least 4 separate studies with a total of at least 35 animals per group. P-values comparing VLP alone to vaccination with VLP and adjuvant are shown, calculated using Fisher's exact tests with stepdown Bonferroni correction. V = VLP, VP = VLP + PolyICLC, VC = VLP + CpG, VM = VLP + MPLA, and VA = VLP + alhydrogel.

Article Snippet: Secondary antibodies included goat anti-mouse IgG-HRP (Southern Biotech 1030–05), IgG1-HRP (Southern Biotech 1070–05), IgG2c-HRP (Southern Biotech 1079–05), and IgG3-HRP (Southern Biotech 1100–05).

Techniques: Enzyme-linked Immunosorbent Assay, Adjuvant

Journal: EBioMedicine

Article Title: Adjuvant-enhanced CD4 T Cell Responses are Critical to Durable Vaccine Immunity

doi: 10.1016/j.ebiom.2015.11.041

Figure Lengend Snippet: Adjuvants have variable impact on IgG subclasses and antibody neutralization. (a) Serum was collected 14 days and 147 days after the vaccine boost (days 35 and 168, respectively) and evaluated for anti-GP IgG, IgG1, IgG2c, and IgG3 levels using an ELISA. Data shown are pooled from at least two separate experiments per group. (b) Pairwise comparison using DSCF multiple pairwise comparison was used and p values greater than 0.05 are shown as “ns”. (c) Summary of results shown in A, B and D. (d) Neutralizing antibody titers were evaluated using the PsVNA, with titers giving 80% neutralization shown; median and IQR shown. Samples within each group were selected randomly from three separate studies for evaluation in the assay. Pairwise comparison using post-hoc Tukey's studentized range test procedure was used to evaluate differences between groups at both days 35 and day 168, where “*” indicates 0.01 < p < 0.05, “**” indicates 0.001 < p < 0.01, “***” indicates 0.0001 < p < 0.001, and “****” indicates p < 0.0001.

Article Snippet: Secondary antibodies included goat anti-mouse IgG-HRP (Southern Biotech 1030–05), IgG1-HRP (Southern Biotech 1070–05), IgG2c-HRP (Southern Biotech 1079–05), and IgG3-HRP (Southern Biotech 1100–05).

Techniques: Neutralization, Enzyme-linked Immunosorbent Assay, Comparison

Journal: EBioMedicine

Article Title: Adjuvant-enhanced CD4 T Cell Responses are Critical to Durable Vaccine Immunity

doi: 10.1016/j.ebiom.2015.11.041

Figure Lengend Snippet: CD8 T cell deficiency does not impact short term or long term survival of vaccinated C57BL/6J mice. (a) Mice were treated IM twice with saline, VLP, VLP and polyICLC, or VLP and CpG. Four weeks after the second vaccination, mice were challenged. Closed symbols represent wild type C57BL/6J mice and open symbols indicate CD8-deficient mice. (b) Mice were vaccinated on the same schedule as in A, but challenge occurred 22 weeks after the second vaccination. (c) Two weeks after the second vaccination and 1 week prior to challenge, blood was collected from vaccinated animals and evaluated for anti-GP IgG antibody titers. (D) A subset of vaccinated animals was euthanized 4 days after the vaccine boost to evaluate CD4 + T cell responses. Median response of C57BL/6J mice and CD8-deficient mice is shown. N = 8–10 per treated group for A–C and D presents data pooled from two separate evaluations of 4–8 mice per group each. “*” indicates 0.005 < p < 0.05.

Article Snippet: Secondary antibodies included goat anti-mouse IgG-HRP (Southern Biotech 1030–05), IgG1-HRP (Southern Biotech 1070–05), IgG2c-HRP (Southern Biotech 1079–05), and IgG3-HRP (Southern Biotech 1100–05).

Techniques: Saline

Journal: Communications Medicine

Article Title: Application of machine learning algorithms to identify serological predictors of COVID-19 severity and outcomes

doi: 10.1038/s43856-024-00658-w

Figure Lengend Snippet: a – d Anti-nucleocapsid (N) and anti-spike (S) secretory IgA or ( e – h ) IgG responses were measured as median fluorescence intensity (MFI) in nasopharyngeal (NP) or oropharyngeal (OP) samples and compared among COVID-19 hospitalized patients classified as moderate (WHO score 3–4), severe (WHO score 5–7), or deceased (WHO score 8). Data are analyzed using Welch’s ANOVA and presented as means with standard deviations, indicated by error bars.

Article Snippet: Plates were incubated for 2 h at RT, washed, and 50 μL of anti-human HRP IgG (1:5000, #A18823, Invitrogen, Thermo Fisher Scientific), IgA (1:5000, #A18787, Invitrogen, Thermo Fisher Scientific), IgG1 (1:4000, #9054-05, Southern Biotech), IgG2 (1:4000, #9060-05, Southern Biotech), IgG3 (1:4000, #9210-05, Southern Biotech) or IgG4 (1:8000, #9200-05, Southern Biotech) secondary antibody was added.

Techniques: Fluorescence

Journal: Communications Medicine

Article Title: Application of machine learning algorithms to identify serological predictors of COVID-19 severity and outcomes

doi: 10.1038/s43856-024-00658-w

Figure Lengend Snippet: a – d IgG binding antibody responses against ancestral spike (S), spike receptor binding domain (S-RBD), and nucleocapsid (N) were quantified by ELISA and calculated as the binding antibody units (BAU) per ml if international standards were available or as the area under the curve (AUC) if standards were not available and titration curves only could be generated; ( e ) ACE2 binding inhibition antibodies were measured using MSD V-PLEX SARS-CoV-2 ACE2 kits; and ( f – h ) Fc effector antibody responses were quantified using complement fixation and antibody-dependent cellular cytotoxicity (ADCC) assays. All assays were run using ancestral SARS-CoV-2. Data were compared using linear regression analysis, controlling for age and biological sex, to look at differences between unvaccinated non-hospitalized and hospitalized patients at 1 MPE. Data are presented as means with standard deviations, indicated by error bars. The limit of detection (LOD) is indicated by the dashed lines. p -values for statistically significant differences (p < 0.05) are shown in the figures.

Article Snippet: Plates were incubated for 2 h at RT, washed, and 50 μL of anti-human HRP IgG (1:5000, #A18823, Invitrogen, Thermo Fisher Scientific), IgA (1:5000, #A18787, Invitrogen, Thermo Fisher Scientific), IgG1 (1:4000, #9054-05, Southern Biotech), IgG2 (1:4000, #9060-05, Southern Biotech), IgG3 (1:4000, #9210-05, Southern Biotech) or IgG4 (1:8000, #9200-05, Southern Biotech) secondary antibody was added.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Titration, Generated, Inhibition

Journal: Communications Medicine

Article Title: Application of machine learning algorithms to identify serological predictors of COVID-19 severity and outcomes

doi: 10.1038/s43856-024-00658-w

Figure Lengend Snippet: The binding ( a – c ) IgG and ( d ) IgA antibodies recognizing ancestral SARS-CoV-2 spike (S), spike receptor binding domain (S-RBD), or nucleocapsid (N) were quantified by ELISA, and measured as the binding antibody units (BAU) per mL if international standards were available or as the area under the curve (AUC) if standards were not available and titration curves could only be generated. e The percentage of ACE2 inhibition for the ancestral SARS-CoV-2 variant was calculated and arcsine transformed for analyses. f – h The Fc effector antibody responses were measured based on C1q complement fixation in response to either the spike or S-RBD or antibody dependent cellular cytotoxicity and reported as arbitrary units (AU). Antibody responses were compared among COVID-19 hospitalized patients classified as moderate (WHO score 3–4), severe (WHO score 5–7), or deceased (WHO score 8) using samples collected at enrollment vs. 1 MPE. Data are presented as means with standard deviations, indicated by error bars. p -values for statistically significant differences (p < 0.05) by linear mixed-effects regression to compare change over time or Welch’s ANOVA to compare across groups within a time point are indicated. Limit of detection (LOD) are indicated by the dashed lines.

Article Snippet: Plates were incubated for 2 h at RT, washed, and 50 μL of anti-human HRP IgG (1:5000, #A18823, Invitrogen, Thermo Fisher Scientific), IgA (1:5000, #A18787, Invitrogen, Thermo Fisher Scientific), IgG1 (1:4000, #9054-05, Southern Biotech), IgG2 (1:4000, #9060-05, Southern Biotech), IgG3 (1:4000, #9210-05, Southern Biotech) or IgG4 (1:8000, #9200-05, Southern Biotech) secondary antibody was added.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Titration, Generated, Inhibition, Variant Assay, Transformation Assay

Journal: Communications Medicine

Article Title: Application of machine learning algorithms to identify serological predictors of COVID-19 severity and outcomes

doi: 10.1038/s43856-024-00658-w

Figure Lengend Snippet: The binding of IgG1 ( a ), IgG2 ( b ), IgG3 ( c ), and IgG4 ( d ) to ancestral SARS-CoV-2 S antigen was measured as the area under the curve (AUC). Spearman correlation of IgG1 ( e ), IgG2 ( f ), IgG3 ( g ), and IgG4 ( h ) with % ACE2 inhibition at enrollment. Hospitalized COVID-19 patients were classified as moderate (WHO score 3–4), severe (WHO score 5–7), or deceased (WHO score 8). Data are presented as means with standard deviations, indicated by error bars. The limit of detection (LOD) is indicated by the dashed lines. p -values for statistically significant differences (p < 0.05) by linear mixed-effects regression to compare change over time or Welch’s ANOVA to compare across groups within a time point are indicated.

Article Snippet: Plates were incubated for 2 h at RT, washed, and 50 μL of anti-human HRP IgG (1:5000, #A18823, Invitrogen, Thermo Fisher Scientific), IgA (1:5000, #A18787, Invitrogen, Thermo Fisher Scientific), IgG1 (1:4000, #9054-05, Southern Biotech), IgG2 (1:4000, #9060-05, Southern Biotech), IgG3 (1:4000, #9210-05, Southern Biotech) or IgG4 (1:8000, #9200-05, Southern Biotech) secondary antibody was added.

Techniques: Binding Assay, Inhibition

Journal: Communications Medicine

Article Title: Application of machine learning algorithms to identify serological predictors of COVID-19 severity and outcomes

doi: 10.1038/s43856-024-00658-w

Figure Lengend Snippet: Linear mixed-effects regression models for ( a – d ) anti-spike (S), anti-spike receptor binding domain (S-RBD), or anti-nucleocapsid (N) IgG or IgA, measured as the binding antibody units (BAU) per ml if international standards were available or as the area under the curve (AUC) if standards were not available and only titration curves could be generated; ( e ) the percentage ACE2 inhibition against ancestral SARS-CoV-2 as a surrogate of virus neutralization, and ( f – h ) Fc effector antibody responses as measured by complement fixation against spike or S-RBD or antibody-dependent cellular cytotoxicity (ADCC) up until 100 DPE or death among hospitalized patients classified as moderate (WHO score 3–4; n = 41), severe (WHO score 5–7; n = 40), or deceased (WHO score 8; n = 24). P -values for statistically significant differences (p < 0.05) by linear mixed-effects regression contrasts are shown within the figures.

Article Snippet: Plates were incubated for 2 h at RT, washed, and 50 μL of anti-human HRP IgG (1:5000, #A18823, Invitrogen, Thermo Fisher Scientific), IgA (1:5000, #A18787, Invitrogen, Thermo Fisher Scientific), IgG1 (1:4000, #9054-05, Southern Biotech), IgG2 (1:4000, #9060-05, Southern Biotech), IgG3 (1:4000, #9210-05, Southern Biotech) or IgG4 (1:8000, #9200-05, Southern Biotech) secondary antibody was added.

Techniques: Binding Assay, Titration, Generated, Inhibition, Virus, Neutralization