Impact of pathogen reduction methods on immunological properties of the COVID‐19 convalescent plasma
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Abstract
COVID‐19 convalescent plasma is an experimental treatment against SARS‐CoV‐2. The aim of this study is to assess the impact of different pathogen reduction methods on the levels and virus neutralizing activity of the specific antibodies against SARS‐CoV2 in convalescent plasma.
Materials and methods
A total of 140 plasma doses collected by plasmapheresis from COVID‐19 convalescent donors were subjected to pathogen reduction by three methods: methylene blue (M)/visible light, riboflavin (R)/UVB and amotosalen (A)/UVA. To conduct a paired comparison, individual plasma doses were divided into 2 samples that were subjected to one of these methods. The titres of SARS‐CoV2 neutralizing antibodies (NtAbs) and levels of specific immunoglobulins to RBD, S‐ and N‐proteins of SARS‐CoV‐2 were measured before and after pathogen reduction.
Results
The methods reduced NtAbs titres differently: among units with the initial titre 80 or above, 81% of units remained unchanged and 19% decreased by one step after methylene blue; 60% were unchanged and 40% decreased by one step after amotosalen; after riboflavin 43% were unchanged and 50% (7%, respectively) had a one‐step (two‐step, respectively) decrease. Paired two‐sample comparisons (M vs. A, M vs. R and A vs. R) revealed that the largest statistically significant decrease in quantity and activity of the specific antibodies resulted from the riboflavin treatment.
Conclusion
Pathogen reduction with methylene blue or with amotosalen provides the greater likelihood of preserving the immunological properties of the COVID‐19 convalescent plasma compared to riboflavin.
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SciScore for 10.1101/2020.10.02.20205567: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Ethical approval was granted for this study by the Independent Moscow City Research Ethical Committee in accordance with national regulations.
Consent: Informed consent was obtained in writing from all donors prior to donation.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After washing the plates 5x the peroxidase-conjugated anti-human IgG detection antibodies diluted in blocking solution were added and plates were incubated at 37°C for 1 h. anti-human IgGsuggested: NoneThe CL-series SARS-CoV-2 IgG and IgM … SciScore for 10.1101/2020.10.02.20205567: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Ethical approval was granted for this study by the Independent Moscow City Research Ethical Committee in accordance with national regulations.
Consent: Informed consent was obtained in writing from all donors prior to donation.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After washing the plates 5x the peroxidase-conjugated anti-human IgG detection antibodies diluted in blocking solution were added and plates were incubated at 37°C for 1 h. anti-human IgGsuggested: NoneThe CL-series SARS-CoV-2 IgG and IgM assays are a two-step chemiluminescent immunoassays for detection of IgG and IgM SARS-CoV-2 antibodies in human serum or plasma, performed on the fully automated Mindray CL 1200i analytical system (Shenzhen Mindray Bio-Medical Electronics Co., Shenzen, China). SARS-CoV-2 IgGsuggested: NoneIgM SARS-CoV-2suggested: NoneAlkaline phosphatase-labeled anti-human IgG or IgM monoclonal antibodies are added to the reaction to form a sandwich with microparticles captured anti-SARS-CoV-2 antibodies. IgMsuggested: Noneanti-SARS-CoV-2suggested: NoneExperimental Models: Cell Lines Sentences Resources Serial 10-fold dilutions of the virus stock were prepared in DMEM with 2% heat- inactivated FBS and in volume of 100 µl were added to Vero E6 cells in a 96-well plate in 8 repeats. Vero E6suggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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