Multistep screening and selection of COVID‐19 convalescent plasma donors at the early stage of the SARS‐CoV‐2 pandemic: A retrospective analysis

Background and Aims The COVID‐19 pandemic reached Bavaria in February 2020. Almost simultaneously, Chinese physicians published reports on the first successful treatments with plasma from COVID‐19 convalescent donors. With these silver linings on the horizon, we decided to establish the manufacturing of anti‐severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) antibody‐containing plasma from COVID‐19 convalescent donors at our site. Here we describe our donor selection process, built from the ground up, which enabled us to cope with the immense resonance after our social media call for donors. Methods As a first step, we created a specific questionnaire for telephone interviews applied by trained students to filter the wave of callers interested in plasma donation. Afterward, the medical staff evaluated the hotline questionnaires and chose eligible donors to be invited for on‐site donor evaluation. Data documentation was performed with MS Excel, and statistical analyses were calculated with GraphPad Prism 8. A quantitative in‐house ELISA was used to detect anti‐SARS‐CoV‐2 antibodies and determine specific titers. Results Out of 1465 calls from potential plasma donors, we could register 420 persons with a completed questionnaire. Evaluation of questionnaires identified 222 of 420 persons as eligible for donation, and 55 were directly asked for on‐site donor qualification. Subsequently, as anti‐SARS‐CoV‐2 antibody titers ≥1:800 were required, we invited 89 of 222 potential donors for an antibody screening. This procedure resulted in another 28 potential donors for an on‐site evaluation. Finally, 12 donors qualified with a titer of 1:400 and 24 with ≥1:800. Conclusion Identifying suitable COVID‐19 convalescent plasma donors was expected to be highly time‐consuming. Implementing a screening procedure with our hotline questionnaire helped us streamline the donor selection process and reduce the workload for the staff. We propose combining the described selection process with the later introduced on‐site antibody screening as an effective strategy.

Multistep screening and selection of COVID-19 convalescent plasma donors at the early stage of the SARS-CoV-2 pandemic: A retrospective analysis.

Background and Aims: The COVID-19 pandemic reached Bavaria in February 2020. Almost simultaneously, Chinese physicians published reports on the first successful treatments with plasma from COVID-19 convalescent donors. With these silver linings on the horizon, we decided to establish the manufacturing of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody-containing plasma from COVID-19 convalescent donors at our site. Here we describe our donor selection process, built from the ground up, which enabled us to cope with the immense resonance after our social media call for donors. Methods: As a first step, we created a specific questionnaire for telephone interviews applied by trained students to filter the wave of callers interested in plasma donation. Afterward, the medical staff evaluated the hotline questionnaires and chose eligible donors to be invited for on-site donor evaluation. Data documentation was performed with MS Excel, and statistical analyses were calculated with GraphPad Prism 8. A quantitative in-house ELISA was used to detect anti-SARS-CoV-2 antibodies and determine specific titers. Results: Out of 1465 calls from potential plasma donors, we could register 420 persons with a completed questionnaire. Evaluation of questionnaires identified 222 of 420 persons as eligible for donation, and 55 were directly asked for on-site donor qualification. Subsequently, as anti-SARS-CoV-2 antibody titers ≥1:800 were required, we invited 89 of 222 potential donors for an antibody screening. This procedure resulted in another 28 potential donors for an on-site evaluation. Finally, 12 donors qualified with a titer of 1:400 and 24 with ≥1:800. Conclusion: Identifying suitable COVID-19 convalescent plasma donors was expected to be highly time-consuming. Implementing a screening procedure with our hotline questionnaire helped us streamline the donor selection process and reduce the workload for the staff. We propose combining the described selection process with the later introduced on-site antibody screening as an effective strategy.

Emergency Use of Convalescent Plasma: Perception of the Regulatory Framework from a Clinical Perspective.

The pandemic spread of an infectious disease poses a plethora of challenges to society, clinicians, health care providers and regulating authorities. In order to mount a rapid response and to provide hope in a potentially catastrophic situation as the current COVID-19 pandemic, emergency plans, regulations and funding strategies have to be developed on regional, national and international levels. The speed needed to establish rapid response programs is challenged by the dynamics of the spread of the disease, the concurrent and competing development of different and potentially more effective treatment options, and not the least by regulatory uncertainty. Convalescent plasma, that is plasma collected from patients who have recovered from COVID-19 infections, has emerged as one of the first potential treatment options in the absence of drugs or vaccines with proven efficacy against SARS-CoV-2. The societal aspects of convalescent plasma and the public awareness gave an additional boost to the rapid employment of convalescent plasma donation platforms immediately after the SARS-CoV-2 outbreak. At the same time, uncertainty remains as to the efficacy of convalescent plasma. With evidence mostly limited to empirical reports, convalescent plasma has been used for decades for the prophylaxis and treatment of various infectious diseases. Clinical trials have addressed different infectious agents, stages of disease, target groups of patients and yielded sometimes inconclusive results. The aim of this short review is to delineate the regulatory background for the emergency use of convalescent plasma in the USA, in the European Union and in Germany, and the transition to the setting of clinical trials. In addition, we describe observations made in the process of collecting COVID-19 convalescent plasma (herein referred to as CCP), and formulate proposals to further improve the framework for rapid responses in future emergency situations.

ECMO in COVID-19-prolonged therapy needed? A retrospective analysis of outcome and prognostic factors.

BACKGROUND: The role of venovenous extracorporeal membrane oxygenation (VV ECMO) in patients with COVID-19-induced acute respiratory distress syndrome (ARDS) still remains unclear. Our aim was to investigate the clinical course and outcome of those patients and to identify factors associated with the need for prolonged ECMO therapy. METHODS: A retrospective single-center study on patients with VV ECMO for COVID-19-associated ARDS was performed. Baseline characteristics, ventilatory and ECMO parameters, and laboratory and virological results were evaluated over time. Six months follow-up was assessed. RESULTS: Eleven of 16 patients (68.8%) survived to 6 months follow-up with four patients requiring short-term (<28 days) and seven requiring prolonged (⩾28 days) ECMO support. Lung compliance before ECMO was higher in the prolonged than in the short-term group (28.1 (28.8-32.1) ml/cmH2O vs 18.7 (17.7-25.0) ml/cmH2O, p = 0.030). Mechanical ventilation before ECMO was longer (19 (16-23) days vs 5 (5-9) days, p = 0.002) and SOFA score was higher (12.0 (10.5-17.0) vs 10.0 (9.0-10.0), p = 0.002) in non-survivors compared to survivors. Low viral load during the first days on ECMO tended to indicate worse outcomes. Seroconversion against SARS-CoV-2 occurred in all patients, but did not affect outcome. CONCLUSIONS: VV ECMO support for COVID-19-induced ARDS is justified if initiated early and at an experienced ECMO center. Prolonged ECMO therapy might be required in those patients. Although no relevant predictive factors for the duration of ECMO support were found, the decision to stop therapy should not be made dependent of the length of ECMO treatment.

Secondary hemophagocytic lymphohistiocytosis and severe liver injury induced by hepatic SARS-CoV-2 infection unmasking Wilson's disease: Balancing immunosuppression.

A 21-year-old woman was hospitalized due to coronavirus disease 2019 (COVID-19)-associated respiratory and hepatic impairment concomitant with severe hemolytic anemia. Upon diagnosis of secondary hemophagocytic lymphohistiocytosis, immunosuppression with anakinra and steroids was started, leading to a hepatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and viremia. Subsequent liver biopsy revealed virus particles in hepatocytes by electron microscopy and SARS-CoV-2 virus could be isolated and cultured. Immunosuppression was stopped and convalescent donor plasma given. In the differential diagnosis, an acute crisis of Wilson's disease was raised by laboratory and genetic testing. This case highlights the complexity of balancing immunosuppression to control hyperinflammation versus systemic SARS-CoV-2 dissemination.

Manufacturing of convalescent plasma of COVID-19 patients: Aspects of quality.

The ongoing coronavirus disease 2019 (COVID-19) pandemic emerged in December 2019. Convalescent plasma represents a promising COVID-19 treatment. Here, we report on the manufacturing of a plasma-based product containing antibodies specific to SARS-CoV-2 obtained from recently recovered COVID-19 patients. Convalescent plasma donors were screened as follows: 1) previously confirmed SARS-CoV-2 infection (by real-time PCR (RT-PCR)); 2) a subsequent negative PCR test followed by a 2-week waiting period; 3) an additional negative PCR test prior to plasmapheresis; and 4) confirmation of the presence of SARS-CoV-2 specific antibodies. Convalescent plasma was stored fresh (2-6°C) for up to 5 days or frozen (-30°C) for long-term storage. Donor peripheral blood and final plasma product were assayed for binding antibodies targeting the SARS-CoV-2 S-protein receptor-binding domain (RBD) and their titers measured by an enzyme-linked immunosorbent assay (ELISA). We performed 72 plasmaphereses resulting in 248 final products. Convalescent plasma contained an RBD-specific antibody titer (IgG) ranging from 1:100 to 1:3200 (median 1:800). The titer was congruent to the titer of the blood (n = 34) before collection (1:100-1:6400, median 1:800). Levels of IL-8 and LBP of donors were slightly increased. Therapeutic products derived from a human origin must undergo rigorous testing to ensure uniform quality and patient safety. Whilst previous publications recommended RBD-specific binding antibody titers of ≥ 1:320, we selected a minimum titer of 1:800 in order to maximize antibody delivery. Production of highly standardized convalescent plasma was safe, feasible and was readily implemented in the treatment of severely ill COVID-19 patients.

A highly specific and sensitive serological assay detects SARS-CoV-2 antibody levels in COVID-19 patients that correlate with neutralization.

OBJECTIVE: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels. METHODS: In our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated. RESULTS: The RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%; N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection; N = 53) in serum. CONCLUSIONS: With the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination.