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. 2022 Oct;14(14):1133-1147.
doi: 10.2217/imt-2022-0015. Epub 2022 Jul 27.

Development and characterization of anti-SARS-CoV-2 intravenous immunoglobulin from COVID-19 convalescent plasma

Mikhail Razumikhin  1 , Tatiana Smolyanova  1 , Alevtina Nikolaeva  2 , Ekaterina Orlova  2 , Aleksandr Ivanov  2 , Olga Belyakova  2 , Tatyana Vyaznikova  2 , Natalia Selezneva  2 , Anton Perevozchikov  2 , Alina Sokolova  2 , Nataliya Zubkova  2 , Irina Efimova  2 , Inna Dolzhikova  3 , Denis Logunov  3 , Elena Sakanjan  2
Affiliations

Development and characterization of anti-SARS-CoV-2 intravenous immunoglobulin from COVID-19 convalescent plasma

Mikhail Razumikhin et al. Immunotherapy. 2022 Oct.

Abstract

Background: The authors describe the developmental process of intravenous anti-COVID-19 hyperimmune immunoglobulin from anti-SARS-CoV-2 neutralizing antibody-containing plasma. Furthermore, the authors investigated its safety and protective activity in animal models. Materials & methods: The manufacturing process included standard ethanol fractionation, chromatographic purification steps and virus removal or inactivation. Results: The authors produced pure and safe immunoglobulin for intravenous administration, with 98.1 ± 6.5 mg/ml protein content, of which 97.6 ± 0.7% was IgG. The concentration factor of SARS-CoV-2 neutralizing antibodies was 9.4 ± 1.4-times. Safety studies in animals showed no signs of acute/chronic toxicity or allergenic or thrombogenic properties. Intravenous anti-COVID-19 hyperimmune immunoglobulin protected immunosuppressed hamsters against SARS-Cov-2. Conclusion: The obtained results can allow the start of clinical trials to study the safety and efficacy in healthy adults.

Keywords: COVID-globulin; IgG; SARS-CoV-2; hyperimmune immunoglobulin; immunoglobulin against COVID-19; preclinical evaluation; protective efficacy.

Plain language summary

An intravenous immunoglobulin with a high concentration of SARS-CoV-2-neutralizing antibodies was prepared from COVID-19 convalescent plasma, which could be utilized as a passive immunization tool in regard to COVID-19 treatment. The manufacturing process employed conforms to commonly held business standards within the intravenous immunoglobulin industry and includes plasma ethanol fractionation following chromatographic purification and special virus removal or inactivation steps. The results of the preclinical in vitro and in vivo experiments demonstrate that the immunoglobulin produced in this study is pure and safe enough to be considered for intravenous applications. The SARS-CoV-2 neutralizing antibody concentration was found to have increased 9.4 ± 1.4-times compared with human plasma. The anti-COVID-19 hyperimmune immunoglobulin showed no signs of toxicity and did not cause any blood clot formations when administered to rabbits. Furthermore, the anti-COVID-19 hyperimmune immunoglobulin was demonstrated to protect immunosuppressed hamsters against SARS-CoV-2.

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Conflict of interest statement

Financial & competing interests disclosure

This work was sponsored and performed by JSC Nacimbio. The employees of Nacimbio, M Razumikhin and T Smolyanova, participated in and contributed to the study. However, the company Nacimbio as a funding source had no direct role in the study design; in the collection, analysis or interpretation of data; in the writing of the report; or in the decision to submit the paper for publication. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

This manuscript was prepared with the help of Editage Publication Support. Writing assistance was sponsored by JSC Nacimbio.

Figures

Figure 1.
Figure 1.. Block flow diagram of the anti-COVID-19 hyperimmune immunoglobulin production process.
HIC: Hydrophobic interaction chromatography; IEC: Ion exchange chromatography.
Figure 2.
Figure 2.. Animal weight and health status.
(A) Change in the body weight of laboratory animals in the different groups after infection. Body weights of the hamsters treated with COVID-globulin, normal IVIG and placebo measured at the indicated days after inoculation with SARS-CoV-2. Group 1 received COVID-globulin 24 h before infection; group 2 received COVID-globulin 2 h after infection; group 3 received COVID-globulin 48 h after infection; group 4 was treated with normal IVIG; group 5 was treated with placebo. Data are presented as mean percentages of the starting weight (± standard error [SE]). *significant difference compared with the IVIG- and placebo-treated groups, p < 0.05; #significant difference in body weight in group 3 compared with that of the normal IVIG- and placebo-treated groups, p < 0.05. (B) Change in the health status of laboratory animals after injection. Health status of animals was assessed on a 5-point scale, where 5: healthy animals; 4: active, reaction to stimuli is normal, flattened ears or tousled coat; 3: weak activity, reaction to stimuli is weak, changes in behavior, aggression or apathy; 2: no response to stimuli, refusal of food and water; 1: death of an animal. Data are presented as the mean average point (± SE). *significant difference compared with the normal IVIG- and placebo-treated groups, p < 0.05; #significant difference in the health status of group 3 compared with that of the normal IVIG- and placebo-treated groups, p < 0.05. COVID-globulin: Anti-COVID-19 hyperimmune immunoglobulin; IVIG: Intravenous immunoglobulin.
Figure 3.
Figure 3.. Survival rate of Syrian hamsters.
Survival was evaluated in animals treated with COVID-globulin, normal intravenous immunoglobulin and placebo at the indicated days after inoculation with SARS-CoV-2. Group 1 received COVID-globulin 24 h before infection; group 2 received COVID-globulin 2 h after infection; group 3 received COVID-globulin 48 h after infection; group 4 was treated with normal intravenous immunoglobulin; group 5 was treated with placebo. COVID-globulin: Anti-COVID-19 hyperimmune immunoglobulin; IVIG: Intravenous immunoglobulin.
Figure 4.
Figure 4.. Median tissue culture infectious dose from lung homogenates and quantification of SARS-CoV-2 RNA copies per 100 mg lung tissue.
The study of viral load was carried out by real-time PCR (A) and using titration in Vero cells (B). Group 1 received COVID-globulin 24 h before infection; group 2 received COVID-globulin 2 h after infection; group 3 received COVID-globulin 48 h after infection; group 4 was treated with normal intravenous immunoglobulin; group 5 was treated with placebo. COVID-globulin: Anti-COVID-19 hyperimmune immunoglobulin; IVIG: Intravenous immunoglobulin.
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