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ORIGINAL RESEARCH
Modified micro test tubes as a promising basis for immobilization of antibodies for immunocapture on the example of SARS-CoV-2
1 Astrakhan State Medical University, Astrakhan, Russia
2 Russian University of Medicine, Moscow, Russia
3 Gabrichevsky Moscow Research Institute for Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Moscow, Russia
4 Gamaleya National Research Centre for Epidemiology and Microbiology, Moscow, Russia
5 Lomonosov Moscow State University, Moscow, Russia
6 Sechenov First Moscow State Medical University, Moscow, Russia
Correspondence should be addressed: Evgeny O. Roubalsky
Bakinskaya, 121, Astrakhan, 414000, Russia; moc.liamg@ykslabur.o.e orp.egahp@ykslabur
Funding: the study was supported by the Russian Science Foundation grant No. 23-15-20035, https://rscf.ru/project/23-15-20035/
Author contribution: Rubalsky EO, Abdrakhmanova RA, Baeva GR, Rubalskaya TS — experimental procedure involving micro test tubes; Shcheblyakov DV, Favorskaya IA — preparation of monoclonal antibodies; Rubalsky EO, Lazko MV, Poroyskiy SV, Gushchin VA — study design. The authors contributed to the search for and analysis of information, preparation of the draft, and manuscript writing equally.
Compliance with ethical standards: the study was approved by the Ethics Committee of the Astrakhan State Medical University. (protocol No. 7 dated 22 February 2023).
The solid-phase immunocapture with antibodies is an important tool used in immunology studies, but conventional polystyrene plates are prone to deformation during thermal cycling and cross-contamination of samples, which reduces accuracy and reproducibility, when molecular genetic testing methods are included in the study. The development of alternative solutions, such as modified polystyrene-coated polypropylene tubes, makes it possible to eliminate these limitations. The study aimed to create a new approach to SARS-CoV-2 immunocapture involving the use of modified test tubes and to assess its efficacy. Monoclonal antibodies P2C5 and R107, as well as inactivated strains GК2020/1 (Wuhan) and hCoV-19/Russia/MOW-PMVL-51/2021 (Omicron) were used for analysis. Immobilization of antibodies, sorption of viral particles, and RNA extraction were accomplished using modified test tubes, standard plates, and uncoated test tubes. The key findings showed that the polystyrene-coated modified test tubes ensured better immunocapture compared to the plates (p < 0.0001), especially when using the P2C5 antibody effective against various SARS-CoV-2 lineages, including Omicron. The R107 antibody showed limited specificity, not exceeding that of the control group with bovine serum albumin. The cross-contamination analysis revealed contamination of 14 samples out of 288 in the plates, while no contamination of samples was reported for modified test tubes. Thus, modified test tubes used for high-precision molecular testing have some advantages, since these decrease the risk of cross-contamination and improve immunocapture efficacy.
Keywords: monoclonal antibodies, SARS-CoV-2, RT-PCR, immunocapture, modified surface, cross-contamination