Page 5 - PULSE- JULY -2020

RGCB becomes India’s first NABL, NABH test strips. We also tested two people who were
and ILAC certified facility approved by Government tested negative through antibody test but had
of India for validation of COVID 19 diagnostics, nasal congestion due to allergy. Both of them
protective gear and anti viral therapeutics passed the Anosmia test showing that general
development. nasal congestion may not severely hamper the
Kit and (iv) Rapid antibody test lateral flow kit Anosmia test using the test strips.
have been approved and is ready for marketing. RGCB initiates program for development of a
The Rapid antigen lateral flow kit, antibody ELISA COVID 19 specific injectable IgG from convalescent Molecular dynamic simulations of RBD-
kit and RT-PCR kit is awaiting final DCGI approval. plasma ACE2 complex for identifying the binding affinities
of SARS-COV-2 RBD mutants.
Research
Studies initiated on pseudovirion
RGCB Covid19 response team investigators endocytosis assays to screen for COVID 19 virus
completed analysis of synonymous mutations in entry inhibitors
S protein of 2939 Covid 19 genomes: COVID-19
pandemic shows considerable variations in disease Transfection of HEK-293T cells with
transmission and pathogenesis globally, yet Lentiviral plasmids (pAX-2; pHIV-EGFP) and SARS
RGCB worked to knock down reasons remain unknown. Our study identifies key CoV-2 S-protein (pCAGGS-Spike) to generate
the lockdown! S-protein mutations prevailing in SARS-CoV-2 strain pseudo-typed virus.
that could alter viral attachment and infectivity.
Over the last few weeks we have all been The investigators propose that the interplay of Harvesting supernatant from HEK-293
overpowered with what is happening all around these mutations could be one of the factors driving cells containing pseudovirions for infection of
the world on Covid-19. The world has changed global variations in COVID-19 spread. In addition, CaCo-2 cells to confirm pseudovirion formation
ever since Covid-19 started unleashing its lethal the mutations identified in this study could be
power. The way we live, work, eat, socialize, etc. an important indicator in predicting efficacies of Studies on HA tagged ACE2 receptor
have changed and have given new dimension vaccines, antibodies and therapeutics that target (pcDNA3.1-ACE2) and FLAG tagged SARS-CoV-2
to our life. Our hearts go out to everyone who SARS-CoV-2 RBD-ACE2 interface (data submitted). S-protein (pCMV14-3X-Flag-SARS-CoV-2 S) in
has been impacted by the virus either directly or progress
indirectly. We also salute those at the front line, Phylogenomic studies in progress to
medical staff and care givers who put themselves understand the evolutionary pattern of SARS-COV-2 Studies on to detect epitope-tagged ACE2
at risk to do all it takes to serve others. circulating in India using 594 high quality SARS- and pseudovirion in progress
COV-2 genomes submitted from 19 states in India.
In much the same way, we at RGCB are doing Cell lines required to generate stable
our bit to fight against this deadly virus using Development of a low cost Anosmia clones of ACE2 for endocytosis assay are ready
our expertise and knowledge gained over the screening tool to mass screen asymptomatic
past three decades to offer solutions and keep COVID-19 carriers: It is known that SARS-CoV-2 A549 cells expressing nuclear targeted m
everyone safe. Towards this, I want to share some first infects the nasopharyngeal region leading to cherry (m Cherry NLS) has been developed to be
of the things done by my very enterprising team at loss of smell. Hence, it was hypothesized that this used for S protein binding assay in 2D and 3D lung
RGCB, over the past few weeks when the country would be the first indicator of COVID-19 infection. models. The cells are to be expressed with human
and RGCB were locked down Therefore, RGCB developed a low cost preliminary ACE2. The expression vector has been designed
screening tool using the initial loss of smell as and cells are ready for transfection.
Developments & Discoveries an indicator that will then need to be followed by
other confirmatory protocols. Covid-19 RT-PCR For the direct binding assay in 2D and 3D
RGCB fulfilled its promise to the Two national collaborations - Nano based positive patients in the age group of 18-60 were models, soluble S protein has been expressed
Honorable Union Minister for Health and Science anti-viral surface coatings for facemasks & PPE subjected to the anosmia test and the results were with GFP fusion in HEK cells. Protein purification
& Technology that it would deliver a complete and use of deep learning for development of very promising. Anosmia was found to be the major is currently ongoing.
COVID 19 Diagnostic suite. Accordingly along with continuous flow type ddPCR. symptom in these patients with 78.72% of them
RGCB’s commercial partner POCT Services, New failing the test. For controls we tested students and A new fluorescent based retroviral
Delhi, the Q LINE series of diagnostic reagents RGCB Director was nominated as member staff returning to RGCB (Antibody negative). All the pseudovirion Assay for SARS-CoV-2: HEK GP
were co-developed. These include (i) Low cost of the PM CARES - COVID 19 Vaccine Expert control subjects passed the Anosmia test using the cells expressing Gag/Pol were stably expressed
Viral Transport Medium, (ii) Viral RNA Extraction Group. strips demonstrating the efficiency of our Anosmia with pBabe puro IMSRP M cherry that express