Production of anti-SARS-CoV-2 hyperimmune globulin from convalescent plasma

AbstractBACKGROUNDIn late 2019, the SARS-CoV-2 virus emerged in China and quickly spread into a world-wide pandemic. Prior to the development of specific drug therapies or a vaccine, more immediately available treatments were sought including convalescent plasma. A potential improvement from convalescent plasma could be the preparation of anti-SARS-CoV-2 hyperimmune globulin (hIVIG).STUDY DESIGN AND METHODSConvalescent plasma was collected from an existing network of plasma donation centers. A caprylate/chromatography purification process was used to manufacture hIVIG. Initial batches of hIVIG were manufactured in a versatile, small-scale facility designed and built to rapidly address emerging infectious diseases.RESULTSProcessing convalescent plasma into hIVIG resulted in a highly purified IgG product with more concentrated neutralizing antibody activity. hIVIG will allow for the administration of greater antibody activity per unit of volume with decreased potential for several adverse events associated with plasma administration. IgG concentration and IgG antibody specific to SARS-CoV-2 were increased over 10-fold from convalescent plasma to the final product. Normalized ELISA activity (per mg/mL IgG) was maintained throughout the process. Protein content in these final product batches was 100% IgG, consisting of 98% monomer and dimer forms. Potentially hazardous proteins (IgM, IgA, and anti-A, anti-B and anti-D antibodies) were reduced to minimal levels.CONCLUSIONSMultiple batches of anti-SARS-CoV-2 hyperimmune globulin (hIVIG) that met regulatory requirements were manufactured from human convalescent plasma. The first clinical study in which the hIVIG will be evaluated will be Inpatient Treatment with Anti-Coronavirus Immunoglobulin (ITAC) [NCT04546581].

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Cross-sectional Analysis of Serum Antibody to Oral Streptococcal Antigens in Children

Journal of Dental Research ◽

10.1177/00220345880670030601 ◽

1988 ◽

Vol 67 (3) ◽

pp. 554-560 ◽

Cited By ~ 13

Author(s):

Z. Luo ◽

D.J. Smith ◽

M.A. Taubman ◽

W.F. King

Keyword(s):

Bacterial Colonization ◽

Serum Antibody ◽

Age Groups ◽

Mutans Streptococci ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Activity ◽

Igg Antibody ◽

Cross Sectional ◽

Streptococcal Species ◽

Micro Organisms

Antibodies to S. salivarius, S. sanguis, and S. mutans cells and to glucosyltransferases (GTF) prepared from these micro-organisms were measured in the sera of 133 infants and children by enzyme-linked immunosorbent assay (ELISA). IgG antibody activity to each cell type and GTF was present at birth (presumably derived from maternal transfer) and declined significantly thereafter. IgG antibody levels to S. salivarius and S. sanguis were next detected in young children (2 to < 3 yr group). However, an increase in IgG antibody to S. mutans cells was not seen until children were older ( 4 to < 8 yr group), possibly reflecting the later colonization of this organism. In contrast, IgG antibody to GTF of all three streptococcal species remained at low levels throughout the first four years of life. IgG antibody to S. mutans GTF was then the first to appear ( 4 to < 8 yr group). Serum IgA antibodies to all GTFs were not detected until after this time. Fifteen sera were used to develop IgG immunoblots with the GTF antigens. Some positive sera (7/12) demonstrated reaction(s) with GTF from each of the three streptococcal species. Individual sera showed IgG antibody bands to GTF from several serotypes of the mutans streptococci. No immunoblot reaction was observed with GTF and sera (3) from the four-to-seven-year and younger age groups. These results indicate the presence of serum antibody to bacteria and bacterial products associated with plaque formation very early in life and during and after the pre-adolescent years. The potential exists for this serum antibody to modulate bacterial colonization or accumulation in the oral cavity.

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Characterization and Implementation of a Diverse Simian Immunodeficiency Virus SIVsm Envelope Panel in the Assessment of Neutralizing Antibody Breadth Elicited in Rhesus Macaques by Multimodal Vaccines Expressing the SIVmac239 Envelope

Journal of Virology ◽

10.1128/jvi.01221-14 ◽

2015 ◽

Vol 89 (16) ◽

pp. 8130-8151 ◽

Cited By ~ 18

Author(s):

Katie M. Kilgore ◽

Megan K. Murphy ◽

Samantha L. Burton ◽

Katherine S. Wetzel ◽

S. Abigail Smith ◽

...

Keyword(s):

Simian Immunodeficiency Virus ◽

Nonhuman Primate ◽

Neutralizing Antibodies ◽

Rhesus Macaques ◽

Neutralizing Antibody ◽

Cd40 Ligand ◽

Antibody Activity ◽

Immunodeficiency Virus ◽

Antibody Profile ◽

Hiv 1

ABSTRACTAntibodies that can neutralize diverse viral strains are likely to be an important component of a protective human immunodeficiency virus type 1 (HIV-1) vaccine. To this end, preclinical simian immunodeficiency virus (SIV)-based nonhuman primate immunization regimens have been designed to evaluate and enhance antibody-mediated protection. However, these trials often rely on a limited selection of SIV strains with extreme neutralization phenotypes to assess vaccine-elicited antibody activity. To mirror the viral panels used to assess HIV-1 antibody breadth, we created and characterized a novel panel of 14 genetically and phenotypically diverse SIVsm envelope (Env) glycoproteins. To assess the utility of this panel, we characterized the neutralizing activity elicited by four SIVmac239 envelope-expressing DNA/modified vaccinia virus Ankara vector- and protein-based vaccination regimens that included the immunomodulatory adjuvants granulocyte-macrophage colony-stimulating factor, Toll-like receptor (TLR) ligands, and CD40 ligand. The SIVsm Env panel exhibited a spectrum of neutralization sensitivity to SIV-infected plasma pools and monoclonal antibodies, allowing categorization into three tiers. Pooled sera from 91 rhesus macaques immunized in the four trials consistently neutralized only the highly sensitive tier 1a SIVsm Envs, regardless of the immunization regimen. The inability of vaccine-mediated antibodies to neutralize the moderately resistant tier 1b and tier 2 SIVsm Envs defined here suggests that those antibodies were directed toward epitopes that are not accessible on most SIVsm Envs. To achieve a broader and more effective neutralization profile in preclinical vaccine studies that is relevant to known features of HIV-1 neutralization, more emphasis should be placed on optimizing the Env immunogen, as the neutralization profile achieved by the addition of adjuvants does not appear to supersede the neutralizing antibody profile determined by the immunogen.IMPORTANCEMany in the HIV/AIDS vaccine field believe that the ability to elicit broadly neutralizing antibodies capable of blocking genetically diverse HIV-1 variants is a critical component of a protective vaccine. Various SIV-based nonhuman primate vaccine studies have investigated ways to improve antibody-mediated protection against a heterologous SIV challenge, including administering adjuvants that might stimulate a greater neutralization breadth. Using a novel SIV neutralization panel and samples from four rhesus macaque vaccine trials designed for cross comparison, we show that different regimens expressing the same SIV envelope immunogen consistently elicit antibodies that neutralize only the very sensitive tier 1a SIV variants. The results argue that the neutralizing antibody profile elicited by a vaccine is primarily determined by the envelope immunogen and is not substantially broadened by including adjuvants, resulting in the conclusion that the envelope immunogen itself should be the primary consideration in efforts to elicit antibodies with greater neutralization breadth.

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The vaccinia-based Sementis Copenhagen Vector COVID-19 vaccine induces broad and durable cellular and humoral immune responses

10.1101/2021.09.06.459206 ◽

2021 ◽

Author(s):

Preethi Eldi ◽

Tamara H Cooper ◽

Natalie A Prow ◽

Liang Liu ◽

Gary K Heinemann ◽

...

Keyword(s):

Immune Responses ◽

Neutralizing Antibodies ◽

First Generation ◽

Neutralizing Antibody ◽

Immune Memory ◽

Antibody Activity ◽

Post Vaccination ◽

Humoral Immune ◽

Humoral Immune Responses ◽

Heterologous Boost

The ongoing COVID-19 pandemic perpetuated by SARS-CoV-2 variants, has highlighted the continued need for broadly protective vaccines that elicit robust and durable protection. Here, the vaccinia virus-based, replication-defective Sementis Copenhagen Vector (SCV) was used to develop a first-generation COVID-19 vaccine encoding the spike glycoprotein (SCV-S). Vaccination of mice rapidly induced polyfunctional CD8 T cells with cytotoxic activity and robust Th1-biased, spike-specific neutralizing antibodies, which are significantly increased following a second vaccination, and contained neutralizing activity against the alpha and beta variants of concern. Longitudinal studies indicated neutralizing antibody activity was maintained up to 9 months post-vaccination in both young and aging mice, with durable immune memory evident even in the presence of pre-existing vector immunity. This immunogenicity profile suggests a potential to expand protection generated by current vaccines in a heterologous boost format, and presents a solid basis for second-generation SCV-based COVID-19 vaccine candidates incorporating additional SARS-CoV-2 immunogens.

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Long-term persistence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein-specific and neutralizing antibodies in recovered COVID-19 patients

10.21203/rs.3.rs-1073046/v1 ◽

2021 ◽

Author(s):

Jira Chansaenroj ◽

Ritthideach Yorsaeng ◽

Nasamon Wanlapakorn ◽

Chintana Chirathaworn ◽

Natthinee Sudhinaraset ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Symptom Onset ◽

Neutralizing Antibodies ◽

Serum Antibody ◽

Neutralizing Antibody ◽

Antibody Responses ◽

Spike Protein ◽

Immunological Study ◽

Igg Antibody ◽

Vaccine Schedule

Abstract Understanding antibody responses after natural severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can guide the coronavirus disease 2019 (COVID-19) vaccine schedule. This study aimed to assess the dynamics of SARS-CoV-2 antibodies, including anti-spike protein 1 (S1) immunoglobulin (Ig)G, anti-receptor-binding domain (RBD) total Ig, anti-S1 IgA, and neutralizing antibody against wild-type SARS-CoV-2 in a cohort of patients who were previously infected with SARS-CoV-2. Between March and May 2020, 531 individuals with virologically confirmed cases of SARS-CoV-2 infection were enrolled in our immunological study. The neutralizing titers against SARS-CoV-2 were detected in 95.2%, 86.7%, 85.0%, and 85.4% of recovered COVID-19 patients at 3, 6, 9, and 12 months after symptom onset, respectively. The seropositivity rate of anti-S1 IgG, anti-RBD total Ig, anti-S1 IgA, and neutralizing titers remained at 68.6%, 89.6%, 77.1%, and 85.4%, respectively, at 12 months after symptom onset. The half-life of neutralizing titers was estimated at 100.7 days (95% confidence interval = 44.5 – 327.4 days, R2 = 0.106). These results support that the decline in serum antibody levels over time depends on the symptom severity, and the individuals with high IgG antibody titers experienced a significantly longer persistence of SARS-CoV-2-specific antibody responses than those with lower titers.

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Immunogenicity of heterologous prime/booster-inactivated and adenoviral-vectored COVID-19 vaccine: real-world data

10.21203/rs.3.rs-785693/v1 ◽

2021 ◽

Author(s):

Nasamon Wanlapakorn ◽

Nungruthai Suntronwong ◽

Harit Phowatthanasathian ◽

Ritthideach Yorsang ◽

Thanunrat Thongmee ◽

...

Keyword(s):

Neutralizing Antibody ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Activity ◽

Serum Samples ◽

Real World Data ◽

Comparison Groups ◽

Chemiluminescent Microparticle Immunoassay ◽

Specific Igg ◽

Vectored Vaccine ◽

Sera Samples

Abstract Limited data are available on the responses to heterologous vaccine regimens for SARS-CoV-2, especially among countries using inactivated and adenoviral-vectored vaccines. A total of 77 participants who received heterologous prime/booster-inactivated COVID-19 vaccine and adenoviral-vectored vaccine were enrolled in our study. There were two comparison groups vaccinated with the homologous CoronaVac (N = 79) and AZD1222 (N = 80) regimen. All sera samples were tested for SARS-CoV-2 spike receptor-binding-domain (RBD) IgG using a chemiluminescent microparticle immunoassay (CMIA). The neutralizing activity in a subset of serum samples was tested against the original Wuhan strain and variants of concern, B.1.1.7 and B.1.351, using an enzyme-linked immunosorbent assay (ELISA)-based surrogate virus neutralization test (sVNT). The CoronaVac followed by the AZD1222 vaccine induced higher levels of spike RBD-specific IgG than that of two-dose CoronaVac or AZD1222 vaccines (p < 0.001). Sera samples of the CoronaVac/AZD1222 vaccine recipients elicited higher neutralizing antibody activity against the original Wuhan and the B.1.351 strain than in the recipients of the two-dose CoronaVac or AZD1222. Following the inactivated CoronaVac/adenoviral-vectored (AZD1222) vaccination administered 14–72 days apart, participants receiving the heterologous vaccine regimen had higher spike RBD-specific IgG and neutralizing activities than the homologous CoronaVac vaccine recipients.

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Immune complexes containing factor V in a patient with an acquired neutralizing antibody

Blood ◽

10.1182/blood.v65.4.810.810 ◽

1985 ◽

Vol 65 (4) ◽

pp. 810-818 ◽

Cited By ~ 20

Author(s):

HC Chiu ◽

AK Rao ◽

C Beckett ◽

RW Colman

Keyword(s):

Immunosorbent Assay ◽

Immune Complexes ◽

Neutralizing Antibody ◽

Circulating Immune Complexes ◽

Heat Inactivation ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration ◽

Factor V ◽

Igg Antibody ◽

Plasma Factor

Abstract An 82-year-old woman presented with extensive hematomas and melena associated with markedly decreased plasma factor V coagulant activity (FV:C). Using a competitive enzyme-linked immunosorbent assay developed in our laboratory, we made serial measurements of factor V antigen (FV:Ag) in plasma and found it to be normal or elevated. The patient's plasma was demonstrated to contain an IgG antibody that could neutralize FV:C in normal plasma. The antibody was of restricted heterogeneity (IgG1, IgG2,kappa). Circulating immune complexes containing antibody to factor V and FV:Ag were demonstrated directly in the plasma by immunoelectrophoresis with polyclonal monospecific antibody and with a monoclonal antibody using an enzyme-linked immunosorbent assay. Presence of neutralizing antibody could be demonstrated in vitro even at times when FV:C was within normal limits by heat inactivation of FV:C. Treatment with plasma and platelet transfusions as well as plasmapheresis induced definite but transient elevation of FV:C. Steroid therapy lowered the neutralizing antibody concentration and produced a rapid and persistent elevation of FV:C during two separate hospitalizations. This report describes a patient in whom levels of FV:Ag have been serially measured, and the presence of circulating immune complexes consisting of factor V and a neutralizing antibody have been directly demonstrated.

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Hemoglobin-Specific Antibody in a Multiply Transfused Patient With Sickle Cell Disease

Blood ◽

10.1182/blood.v89.6.2155 ◽

1997 ◽

Vol 89 (6) ◽

pp. 2155-2158 ◽

Cited By ~ 5

Author(s):

Peter A. Noronha ◽

Loyda N. Vida ◽

C. Lucy Park ◽

George R. Honig

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Solid Phase ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Activity ◽

Cell Disease ◽

Serum Samples ◽

Igg Antibody ◽

Microtiter Plates ◽

Level Of Activity

Abstract Human hemoglobins (Hbs) are known to be immunogenic, and both normal and variant forms of Hb have been shown to stimulate antibody formation in a variety of animal species. In patients who are homozygous for the sickle Hb (HbS) mutation, transfusion of normal, HbA-containing erythrocytes provides a potential stimulus for HbA alloimmunization. We tested serum samples for the presence of anti-Hb antibody by a solid-phase enzyme-linked immunosorbent assay (ELISA) using Hb-coated polystyrene microtiter plates. Hb-bound antibody was identified using an antihuman IgG antibody. Serum samples from 89 patients with sickle cell disease were initially tested for evidence of Hb antibody. The serum from three individuals exhibited antibody activity against HbA with little or no activity against HbS. Only one of them, a multiply transfused adult with HbSS, was available for further study. When this patient's antibody was tested against a variety of normal and mutant Hbs using antibody either to human IgG or to κ chains, the anti-Hb antibody demonstrated specificity for the region of the Hb β chain corresponding to the site of the amino acid substitution of HbS. The level of activity of the patient's anti-HbA showed no significant change over 1.5 years of observation. The transfusion of erythrocytes containing Hb structurally different from that of the recipient appeared to be capable of stimulating the production of Hb-specific alloimmune antibody.

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Multicenter Evaluation of the Clinical Performance and the Neutralizing Antibody Activity Prediction Properties of ten high throughput serological assays used in Clinical Laboratories.

Journal of Clinical Microbiology ◽

10.1128/jcm.02511-20 ◽

2020 ◽

Author(s):

C Therrien ◽

B Serhir ◽

M Bélanger-Collard ◽

J Skrzypczak ◽

D. K. Shank ◽

...

Keyword(s):

Clinical Performance ◽

Neutralizing Antibody ◽

High Specificity ◽

Roc Curves ◽

Antibody Activity ◽

Activity Prediction ◽

Predictive Values ◽

Population Immunity ◽

Accurate Comparison ◽

Diagnostic Resolution

As the COVID-19 pandemic second wave is emerging, it is of the upmost importance to screen the population immunity in order to keep track of infected individuals. Consequently, SARS-CoV-2 immunoassays with high specificity and positive predictive values are needed to obtain an accurate epidemiological picture. As more data accumulate about the immune responses and the kinetics of neutralizing antibody (nAb) production in SARS-CoV-2 infected individuals, new applications are forecasted for serological assays such as nAb activity prediction in convalescent plasma from recovered patients. This multicenter study, involving six hospital centres, determined the baseline clinical performances, reproducibility and nAb level correlations of ten commercially available immunoassays. In addition, three lateral flow chromatography assays were evaluated as these devices can be used in logistically challenged area. All assays were evaluated using the same patient panels in duplicate thus enabling accurate comparison of the tests. Seven immunoassays examined in this study were shown to have excellent specificity (98 to 100%) and good to excellent positive predictive values (82 to 100%) when used in a low (5%) seroprevalence setting. We observed sensitivity values as low as 74% and as high as 95% at ≥15 days post symptom onset. The determination of optimized cut-off values through ROC curves analyses had a significant impact on the diagnostic resolution of several enzyme immunoassays by increasing the sensitivity significantly without a large trade-off in specificity. We found that Spike-based immunoassays seems to be better correlates of nAb activity. Finally, the results reported here will add up to the general knowledge of the inter-laboratory reproducibility of clinical performance parameters of immunoassays and provide new evidence about nAb activity prediction.

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A randomized comparison of two doses of human lymphoblastoid interferon- alpha in hairy cell leukemia. Wellcome HCL Study Group

Blood ◽

10.1182/blood.v78.12.3133.3133 ◽

1991 ◽

Vol 78 (12) ◽

pp. 3133-3141 ◽

Cited By ~ 1

Author(s):

RV Smalley ◽

SA Anderson ◽

RL Tuttle ◽

J Connors ◽

LM Thurmond ◽

...

Keyword(s):

Platelet Count ◽

Beneficial Effect ◽

Hairy Cell Leukemia ◽

Low Dose ◽

Neutralizing Antibody ◽

Chronic Fatigue ◽

Initial Therapy ◽

Antibody Activity ◽

Hairy Cell ◽

Cell Leukemia

Abstract One hundred thirty-eight patients with hairy cell leukemia were randomized to receive either a dose of 2.0 megaunits (MU)/m2 or a 10- fold lower dose of 0.2 MU/m2 of a highly purified natural alpha- interferon, administered daily for 28 days followed by a three times a week schedule. Ninety-seven of these patients had previously undergone splenectomy, but otherwise none of the patients had received prior therapy for their leukemia. The two doses were comparable in their effect on improving the neutrophil and platelet count, whereas the higher dose had a greater beneficial effect on the hemoglobin level and a greater antileukemic effect on the marrow. Acute toxicity in the form of a flu-like syndrome, neurologic side effects, neutropenia, and the need for platelet transfusions was observed less frequently in the low- dose group, as was the chronic fatigue syndrome. No neutralizing antibody activity was seen in the sera from 61 patients examined. Because of its beneficial effect on the neutrophil and platelet count and a lower degree of toxicity (ie, a superior therapeutic/toxicity ratio), the low dose is recommended as initial therapy in patients with hairy cell leukemia. This therapy may be followed by dose escalation once clinical improvement is observed.

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