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Updated: Nov 13, 2025

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection
Published on: June 5, 2021
Development of neutralizing antibody responses against SARS-CoV-2 in COVID-19 patients
María Teresa Valenzuela1, Cinthya Urquidi1, Nicolás Rodriguez2
1Department of Epidemiology and Public Health, Faculty Medicina, Universidad de los Andes, Santiago, Chile.
Abstract:
The novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19) are new global problems. The understanding of the host immune response in COVID-19 and its implications in the development of therapeutic agents are new challenges. Here, we evaluated the development of immunoglobulin G (IgG) and neutralizing (Nt) antibodies in symptomatic hospitalized COVID-19 patients. We followed up 117 COVID-19 confirmed patients from a reference health center for COVID-19 during the epidemic in Santiago de Chile. One and two sequential blood samples from 117 to 68 cases were, respectively, obtained to evaluate the immune response. Immunofluorescence and neutralization assays in Vero E6 cells with a Chilean SARS-CoV-2 strain were performed. Out of the 68 patients, 44% were women and 56% men, and the most frequent comorbidities were hypertension (47.7%) and diabetes (27.4%). The most frequent symptoms or signs related to COVID-19 were dyspnea, cough, fever, myalgia, and headache. In all the study population, 76.1% and 60.7% of patients were positive for IgG and Nt antibodies in the first blood sample. All patients except one were positive for IgG and Nt antibodies in the second sample. IgG and Nt antibodies positivity increased significantly according to the disease evolution periods. Higher Nt antibody titers were observed in the first sample in patients under 60 years of age. Obese and diabetic patients had no increase in Nt antibodies, unlike normal weight and diabetes-free patients. Both hypertensive and normotensive patients showed a significant increase in Nt antibodies. These results show an early and robust immune response against SARS-CoV-2 infection during severe COVID-19.
Insights
This study shows that hospitalized COVID-19 patients develop robust immunoglobulin G (IgG) and neutralizing (Nt) antibodies early in their infection. Antibody levels significantly increase over time, indicating an effective immune response to SARS-CoV-2.
Area of Science:
- Immunology
- Virology
- Infectious Diseases
Background:
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), a significant global health challenge.
- Understanding the host immune response is crucial for developing effective COVID-19 therapeutics.
- The development of specific antibodies, including immunoglobulin G (IgG) and neutralizing (Nt) antibodies, is a key aspect of this response.
Purpose of the Study:
- To evaluate the development and kinetics of IgG and Nt antibodies in symptomatic, hospitalized COVID-19 patients.
- To assess the correlation between antibody response and patient demographics, comorbidities, and disease progression.
Main Methods:
- Longitudinal study of 117 hospitalized COVID-19 patients in Santiago, Chile.
- Collection of sequential blood samples for antibody analysis.
- Detection of IgG and Nt antibodies using immunofluorescence and neutralization assays against a local SARS-CoV-2 strain.
Main Results:
- In the first blood sample, 76.1% of patients had IgG and 60.7% had Nt antibodies.
- Antibody positivity increased significantly by the second sample, with all but one patient testing positive.
- Higher Nt antibody titers were observed in younger patients (<60 years).
- Obese and diabetic patients showed diminished Nt antibody increases compared to normal weight and non-diabetic individuals.
- Both hypertensive and normotensive patients demonstrated a significant increase in Nt antibodies.
Conclusions:
- Symptomatic, hospitalized COVID-19 patients exhibit an early and robust antibody response to SARS-CoV-2.
- Antibody levels increase significantly with disease evolution.
- Factors such as age, obesity, and diabetes may influence the neutralizing antibody response.
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