Neutralizing activity to SARS‐CoV‐2 of convalescent and control plasma used in a randomized controlled trial

Freedenberg, Alex T.; Pan, Chun‐Hao; Diehl, William E.; Romeiser, Jamie L.; Hwang, Ga‐Ram; Leiton, Cindy V.; Muecksch, Frauke; Shroyer, Kenneth R.; Bennett‐Guerrero, Elliott

BACKGROUND: There are limited data on the neutralizing activity of convalescent plasma (CP) administered in randomized controlled trials (RCT) of COVID‐19 infection. STUDY DESIGN AND METHODS: As part of an RCT, CP was collected per FDA guidelines from individuals recovered from COVID‐19 infection. CP donors had to have ≥145 optical density (OD) units (ideal target ≥300) using a semiquantitative, immunochromatographic test for IgG antibody to the nucleocapsid protein (NP) of SARS‐CoV‐2 (typical range 0–500 OD units). A random subset of samples [14 control plasma, 12 CP “medium‐anti‐NP” (145–299 OD units), and 13 CP “high” anti‐NP (≥300 OD units)] were tested for neutralizing antibodies using an established viral luciferase antibody inhibition assay to detect the infection of SARS‐CoV‐2 pseudovirus that encoded spike protein (SARS2‐S(trunc)) on a human immunodeficiency virus 1 vector (NL43dEnvNanoLuc), using ACE2‐expressing 293 T cells. The titer needed to neutralize 50% of viral activity (NT50) was calculated. RESULTS: The uptake of SARS‐CoV‐2 pseudovirus by 293T(ACE2) cells was inhibited by pretreatment with CP compared to control CP (p < .001) with control plasma having a median (IQR) 50% neutralization titer (NT50) of 1:28 (1:16,1:36) compared to 1:334 (1:130,1:1295) and 1:324 (1:244,1:578), for medium anti‐NP and high anti‐NP CP units, respectively. The neutralizing activity of CP met minimum FDA criteria with neutralizing antibody titers >1:80 in 100% of randomly selected samples, using a conservative approach that excluded non‐specific binding. DISCUSSION: Plasma from donors screened using an immunochromatographic test for IgG antibody to SARS‐CoV‐2 NP exhibited neutralizing activity meeting FDA's minimum standard in all randomly selected COVID‐19 CP units.