Production of hyperimmune anti-SARS-CoV-2 intravenous immunoglobulin from pooled COVID-19 convalescent plasma

Shaukat Ali, Syed M Uddin, Ayesha Ali, Fatima Anjum, Rashid Ali, Elisha Shalim, Mujtaba Khan, Iqra Ahmed, Sheikh M Muhaymin, Uzma Bukhari, Shobha Luxmi, Abdul S Khan, Saeed Quraishy, Shaukat Ali, Syed M Uddin, Ayesha Ali, Fatima Anjum, Rashid Ali, Elisha Shalim, Mujtaba Khan, Iqra Ahmed, Sheikh M Muhaymin, Uzma Bukhari, Shobha Luxmi, Abdul S Khan, Saeed Quraishy

Abstract

Background: This study assesses the feasibility of producing hyperimmune anti-COVID-19 intravenously administrable immunoglobulin (C-IVIG) from pooled convalescent plasma (PCP) to provide a safe and effective passive immunization treatment option for COVID-19. Materials & methods: PCP was fractionated by modified caprylic acid precipitation followed by ultrafiltration/diafiltration to produce hyperimmune C-IVIG. Results: In C-IVIG, the mean SARS-CoV-2 antibody level was found to be threefold (104 ± 30 cut-off index) that of the PCP (36 ± 8.5 cut-off index) and mean protein concentration was found to be 46 ± 3.7 g/l, comprised of 89.5% immunoglobulins. Conclusion: The current method of producing C-IVIG is feasible as it uses locally available PCP and simpler technology and yields a high titer of SARS-CoV-2 antibody. The safety and efficacy of C-IVIG will be evaluated in a registered clinical trial (NCT04521309).

Keywords: SARS-CoV-2; anti-COVID-19 IVIG; caprylic acid; convalescent plasma; diafiltration; immunotherapy; intravenous immunoglobulins; passive immunization; pooled plasma; ultrafiltration.

Figures

Figure 1.. Process flow diagram for the…
Figure 1.. Process flow diagram for the production of high-titer anti-SARS-CoV-2 intravenously administrable immunoglobulin.
IVIG: Intravenously administrable immunoglobulin.
Figure 2.. Immunoglobulin content and anti-SARS-CoV-2 antibody…
Figure 2.. Immunoglobulin content and anti-SARS-CoV-2 antibody level.
(A) Comparison of mean IgG, IgM, IgA and anti-SARS-CoV-2 antibody levels in pooled convalescent plasma with that in C-IVIG and control IVIG produced by the caprylic acid precipitation method. Control IVIG (pre-pandemic) was found to be nonreactive for SARS-CoV-2 antibodies. The anti-SARS-CoV-2 antibody level is measured as cut-off index (sample absorbance/cut-off; cut-off = 1). (B) Comparison of IgG, IgM and IgA content (as percentage of total immunoglobulin) in pooled plasma, C-IVIG and control IVIG. (C) Mean percentage yield of IgG, IgA, IgM and anti-SARS-CoV-2 antibodies from convalescent plasma fractionation. All values are represented as the mean of eight consecutive batches of C-IVIG production. C-IVIG: Anti-COVID-19 intravenously administrable immunoglobulin; IVIG: Intravenously administrable immunoglobulin.
Figure 3.. SDS-PAGE.
Figure 3.. SDS-PAGE.
Patterns of eight consecutive batches of IVIG in (A) non-reducing and (B) reducing conditions. B1–B8: Batch numbers; IVIG: Intravenously administrable immunoglobulin; Mw.M: Molecular weight markers (10–250 KDa).
Figure 4.. Histological examination of liver and…
Figure 4.. Histological examination of liver and kidney of rats.
(A) Cross-section of kidney treated with 0.5 mg/kg dose of intravenously administrable immunoglobulin. (B) Cross-section of liver treated with 0.5 mg/kg dose. (C) Cross-section of kidney of control group. (D) Cross-section of liver of control group.

References

    1. Wang D, Hu B, Hu C et al. Clinical characteristics of 138 hospitalized patients with 2019 novel coronavirus-infected pneumonia in Wuhan, China. JAMA 323(11), 1061–1069 (2020).
    1. Huang C, Wang Y, Li X et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 395(10223), 497–506 (2020).
    1. Rothan HA, Byrareddy SN The epidemiology and pathogenesis of coronavirus disease (COVID-19) outbreak. J. Autoimmun. 109, 102433 2020).
    1. World Health Organization. Coronavirus disease (COVID-19) pandemic (2020).
    1. Fernandes N Economic effects of coronavirus outbreak (COVID-19) on the world economy. IESE Business School Working Paper No. WP-1240-E. (2020) (Epub ahead of print)
    1. Cunningham AC, Goh HP, Koh D Treatment of COVID-19: old tricks for new challenges. Critical Care 24(1), 91 (2020).
    1. Kim JH, Marks F, Clemens JD. Looking beyond COVID-19 vaccine phase 3 trials. Nat. Med. 1–7 (2021).
    1. Chen L, Xiong J, Bao L, Shi Y Convalescent plasma as a potential therapy for COVID-19. Lancet Infect. Dis. 20(4), 398–400 (2020).

    2. • Supported the concept of our study.

    1. Tanne JH Covid-19: FDA approves use of convalescent plasma to treat critically ill patients. BMJ 368, m1256 2020).

    2. • Supported the concept of our study.

    1. Shen C, Wang Z, Zhao F et al. Treatment of 5 critically ill patients with COVID-19 with convalescent plasma. JAMA 323(16), 1582–1589 (2020).

    2. • Supported the concept of our study.

    1. Chai KL, Valk SJ, Piechotta V et al. Convalescent plasma or hyperimmune immunoglobulin for people with COVID‐19: a living systematic review. Cochrane Database Syst.Rev. 10, CD013600 2020).
    1. Cao W, Liu X, Bai T et al. High-dose intravenous immunoglobulin as a therapeutic option for deteriorating patients with coronavirus disease 2019. Open Forum Infect. Dis. 7(3),ofaa102 (2020).
    1. Xie Y, Cao S, Li Q et al. Effect of regular intravenous immunoglobulin therapy on prognosis of severe pneumonia in patients with COVID-19. J. Infect. 81(2), 318–356 2020).
    1. Salazar G, Zhang N, Fu TM et al. Antibody therapies for the prevention and treatment of viral infections. NPJ Vaccines 2, 19 (2017).
    1. De Alwis R, Chen S, Gan ES, Ooi EE Impact of immune enhancement on Covid-19 polyclonal hyperimmune globulin therapy and vaccine development. EBioMedicine 55, 102768 2020).

    2. • Clarified IVIG’s potential mode of action in COVID-19.

    1. León G et al. Industrial Production and Quality Control of Snake Antivenoms. : Toxinology. Gopalakrishnakone P, Calvete J (). Springer, Dordrecht: (2014).
    1. El-Ekiaby M, Vargas M, Sayed M et al. Minipool caprylic acid fractionation of plasma using disposable equipment: a practical method to enhance immunoglobulin supply in developing countries. PLoS Negl. Trop. Dis. 9(2), (2015).

    2. • Provided evidence on human plasma fractionation through caprylic acid precipitation.

    1. Morais V, Berasain P, Massaldi H Immunoglobulin purification by caprylic acid. Protein Downstream Processing. Humana Press, NJ, USA: 137–143 (2014).

    2. • Provided a standard to compare the process efficiency.

    1. Laemmli, U. K Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227(5259), 680–685 (1970).
    1. U.S. Pharmacopeia. USP-71,Sterility tests. United States Pharmacopeia and National Formulary. USP 41-NF 36 U.S. Pharmacopeial Convention, MD, USA, 5984–5991 (2018).
    1. Pharmacopeia 41 U.S. 151 Pyrogen Test. United States Pharmacopeial Convention, Inc. MD, USA,6083–6085 (2018).
    1. Keller MA, Stiehm ER Passive immunity in prevention and treatment of infectious diseases. Clin. Microbiol. Rev. 13(4), 602–614 (2000).
    1. Duan K, Liu B, Li C et al. Effectiveness of convalescent plasma therapy in severe COVID-19 patients. Proc. Natl Acad. Sci. USA 117(17), 9490–9496 (2020).
    1. Abrams-Downey A, Saabiye J, Vidaurrazaga M Investigational therapies for the treatment of COVID-19: updates from ongoing clinical trials. Eur. Urol. Focus 6(5), 1028–1031 (2020).
    1. Nguyen AA, Habiballah SB, Platt CD, Geha RS, Chou JS, Mcdonald DR Immunoglobulins in the treatment of COVID-19 infection: proceed with caution! Clin. Immunol. 216, 108459 2020).

    2. • Provided evidence on the possible role of hyperimmune immunoglobulin therapy for COVID-19.

    1. Weidner L, Gänsdorfer S, Unterweger S et al. Quantification of SARS-CoV-2 antibodies with eight commercially available immunoassays. J. Clin. Virol. 129, 104540,2020).

    2. • Provided authentication of the Elecsys system used to quantify anti-SARS-CoV-2 antibodies.

    1. Barahona Afonso AF, João CMP The production processes and biological effects of intravenous immunoglobulin. Biomolecules 6(1), 15 (2016).
    1. Harlow ED, Lane D Antibodies: a Laboratory Manual. Cold Spring Harbor Laboratory, NY, USA, 579 (1988).
    1. Alberts B, Johnson A, Lewis J, Morgan D, Raff M, Keith Roberts PW W. W. Norton & Company; Molecular Biology of the Cell NY, USA: . (2018).
    1. Fundenberg HH, Stites DP, Caldwell JL, Wells JV Basic and Clinical Immunology. Lange Medical Publications, CA, USA: (1976).
    1. Radosevich M, Burnouf T Intravenous immunoglobulin G: trends in production methods, quality control and quality assurance. Vox. Sang. 98(1), 12–28 (2010).
    1. Wang Z, Lorenzi JC, Muecksch F et al. Enhanced SARS-CoV-2 neutralization by secretory IgA in vitro. bioRxiv 2020) (Epub ahead of print).
    1. Parkkinen J, Rahola A, Von Bonsdorff L, Tölö H, Törmä E A modified caprylic acid method for manufacturing immunoglobulin G from human plasma with high yield and efficient virus clearance. Vox. Sang. 90(2), 97–104 (2006).
    1. Kothe N, Rudnick D, Piechaczek D, Klein H, Rohm D, Kloft M (Manufacturing intravenous tolerable immunoglobulin-g preparation US5164487A United States: 1992).
    1. Vargas M, Segura A, Wu YW et al. Human plasma‐derived immunoglobulin G fractionated by an aqueous two‐phase system, caprylic acid precipitation, and membrane chromatography has a high purity level and is free of detectable in vitro thrombogenic activity. Vox. Sang. 108(2), 169–177 (2015).
    1. Yang R, Lan J, Huang B et al. Lack of antibody-mediated cross-protection between SARS-CoV-2 and SARS-CoV infections. EBioMedicine 58, 102890 2020).
    1. Abolhassani H, Asgardoon MH, Rezaei N, Hammarstrom L, Aghamohammadi A Different brands of intravenous immunoglobulin for primary immunodeficiencies: how to choose the best option for the patient?. Expert. Rev. Clin. Immunol. 11(11), 1229–1243 (2015).
    1. Ochiai M, Yamamoto A, Naito S et al. Applicability of bacterial endotoxins test to various blood products by the use of endotoxin-specific lysates. Biologicals 38(6), 629–636 (2010).

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