Journal: bioRxiv

Article Title: Inhibitory activities of monoclonal antibodies against Staphylococcus aureus Clumping factor A

doi: 10.1101/2025.05.21.655433

Figure Lengend Snippet: (A) Domain organization of ClfA depicting the signal sequence (S), N1, N2, N3 subdomains, A domain (ClfA-A), serine aspartic repeat region (SDR), LPXTG motif for attachment to peptidoglycan by sortase A (black rectangle). N2-N3 segment involved in fibrinogen (Fg)/fibrin binding and binding sites of mAbs deduced from this study are shown. (B, C) Coomassie stained gels of purified mAbs (non-reducing conditions) (B) and ClfA domains and subdomains from various strains (C). Numbers to the left of gels indicate molecular weight markers in kDa. (D-H) Representative ELISA showing mAb binding to ClfA-A Newman, N315, WU1, USA300 as well as isolated, ClfA-N1, -N2 or -N3 subdomains from strain Newman. Antibodies are presented as follows: 2F10 (D), 8B9 (E), 1A5 (F), 1C2 (G), 2A12 (H). Bound antibodies were detected with polyclonal anti-mouse HRP-conjugated secondary antibody and reported as absorbance at 450 nm (A 450 ). Association constants and comparative analyses are shown in and Supplementary Table 1. (I) Relative abundance of Newman ClfA-A, N1, N2 and N3 antibodies in IVIG. Data was analyzed with GraphPad Prism 10 using non-linear fit (least squared) regression. (**, P < 0.01; ns: not significant). All experiments were performed at least twice.

Article Snippet: To evaluate the presence of antibodies against ClfA in human sera, serial dilutions of IVIG (Privigen® 10% Liquid Intravenous Immunoglobulin, CSL Behring) were prepared by an initial dilution of 1:10,000 in blocking buffer, followed by seven 1:5 serial dilutions.

Techniques: Sequencing, Binding Assay, Staining, Purification, Molecular Weight, Enzyme-linked Immunosorbent Assay, Isolation

Journal: medRxiv

Article Title: Intravenous Immunoglobulin (IVIG) in Treating Non-ventilated COVID-19 Patients with Moderate to Severe Hypoxia is Pharmacoeconomically Favorable When Appropriately Targeted

doi: 10.1101/2021.09.26.21264152

Figure Lengend Snippet: Costs of IVIG and control COVID-19 cases with respect to average reimbursement by Medicaid (solid line, bottom), Medicare (dashed line, middle), and commercial insurance (dotted line, top) obtained from reference 12.

Article Snippet: IVIG (Privigen 10%, CSL Behring) was utilized off-label to treat moderate to severely ill non-ICU patients with COVID-19 requiring > 3 liters O2 by nasal cannula, not mechanically ventilated, but considered at high risk for respiratory failure.

Techniques:

Journal: Thrombosis Research

Article Title: COVID-19 adenovirus vaccine triggers antibodies against PF4 complexes to activate complement and platelets

doi: 10.1016/j.thromres.2021.10.027

Figure Lengend Snippet: Characteristics of the patients. Anti-PF4 antibodies ELISA (Asserachrom HPIA, Diagnostica Stago) with global recognition of immunoglobulins. Interpretation of ELISA result, <30% of reference; negative, 30–50%, borderline positive, >50% positive, >100% strongly positive. IVIG; intravenous immunoglobulin (dose see p. 7–9). Reference values for hemoglobin (male 134–167 g/L and for female 117–155 g/L), platelet count (150–360 × 10 9 /L), PT; prothrombin time (70–130% of normal), fibrinogen (2.0–4.0 g/L) and D-dimer ( <0.5 mg/L).

Article Snippet: The effects of human immunoglobulin (IVIG) (5 and 10 mg/mL; Privigen® CSL Behring, Marburg, Germany, the brand for patient treatment), glycoprotein (GP) IIb/IIIa inhibitor eptifibatide (2 μg/mL; Integrelin®, GSK, Dublin, Ireland) and recombinant human PF4 (ab9562, Abcam B.V., Amsterdam, Netherlands) on platelet aggregation were assessed.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Thrombosis Research

Article Title: COVID-19 adenovirus vaccine triggers antibodies against PF4 complexes to activate complement and platelets

doi: 10.1016/j.thromres.2021.10.027

Figure Lengend Snippet: A-C. Spontaneous (A, C) and collagen-induced healthy donor platelet aggregation (B, C) triggered by the acute phase plasma from patient 1. (A) Spiking with patient EDTA-plasma: spontaneous aggregation (no agonist) of platelets from healthy donor (duplicate). Glycoprotein (GP) IIb/IIIa inhibitor (eptifibatide) (2 μg/mL) abolished the spontaneous aggregation, as did intravenous immunoglobin (IVIG) (5 mg/mL). (B) Spiking with control plasma: spontaneous aggregation (no agonist) did not occur. Collagen-induced aggregation supplemented with platelet factor 4 (PF4, 4 μg/mL) was not enhanced. (C) Spiking with patient plasma: immediate spontaneous aggregation in platelets spiked with patient plasma occurred, and continued upon adding collagen. Addition of PF4 enhanced collagen-induced aggregation. GPIIb/IIIa inhibitor and IVIG at 10 mg/mL both inhibited collagen-induced aggregation, but IVIG at 5 mg/mL had only a moderate response with control (B) or patient plasma (C). PF4 did not counteract the inhibitory effect of GPIIb/IIIa inhibitor or IVIG (10 mg/mL) in control (B) or patient plasma (C).

Article Snippet: The effects of human immunoglobulin (IVIG) (5 and 10 mg/mL; Privigen® CSL Behring, Marburg, Germany, the brand for patient treatment), glycoprotein (GP) IIb/IIIa inhibitor eptifibatide (2 μg/mL; Integrelin®, GSK, Dublin, Ireland) and recombinant human PF4 (ab9562, Abcam B.V., Amsterdam, Netherlands) on platelet aggregation were assessed.

Techniques:

Journal: Thrombosis Research

Article Title: COVID-19 adenovirus vaccine triggers antibodies against PF4 complexes to activate complement and platelets

doi: 10.1016/j.thromres.2021.10.027

Figure Lengend Snippet: A-F. Spontaneous and collagen-induced healthy donor platelet aggregation triggered by the acute (A, B) versus follow-up phase plasma (C, D) and serum (E, F) from patient 3. (A) Spiking with control plasma without agonist and in collagen-induced aggregation: spontaneous aggregation did not occur (no agonist) and collagen-induced aggregation was vivid. (B) Spiking with patient plasma at acute phase induced immediate spontaneous aggregation (no agonist), enhanced by collagen. Collagen-induced platelet aggregation was modestly inhibited by intravenous immunoglobin (IVIG) (5 mg/mL) when spiked with control plasma (A) or on patient plasma (B), while Glycoprotein (GP) IIb/IIIa inhibitor (eptifibatide) (2 μg/mL) abolished aggregation. The five-week follow-up patient plasma did not trigger spontaneous aggregation (D), but patient serum depicted delayed spontaneous aggregation (F). Compared with plasma (C) or serum controls (E), collagen-induced aggregation in the presence of patient plasma (D) or serum (F) was nearly normal, despite ticagrelor and fondaparinux medication. Adding platelet factor 4 (PF4) (5 μg/mL) did not markedly enhance aggregation (C-F), but IVIG 10 mg/mL abolished aggregation in the presence of PF4 (C-F).

Article Snippet: The effects of human immunoglobulin (IVIG) (5 and 10 mg/mL; Privigen® CSL Behring, Marburg, Germany, the brand for patient treatment), glycoprotein (GP) IIb/IIIa inhibitor eptifibatide (2 μg/mL; Integrelin®, GSK, Dublin, Ireland) and recombinant human PF4 (ab9562, Abcam B.V., Amsterdam, Netherlands) on platelet aggregation were assessed.

Techniques: