ADVERTORIAL: Premas Biotech’s D-crypt platform

Premas Biotech’s D-cryptTM platform for Virus Like Particles (VLPs) based vaccine candidates against SARS-CoV-2

It’s been more than eighteen months, and we are still conquering COVID-19. Previously the first wave and now the second wave has become a nightmare for human beings. India is following the US in terms of cases of COVID-19 infections. During the lethal second wave of COVID-19 infections, India has recorded nearly more than 2.5 lakh deaths which are way higher than fatalities caused during the first wave. The mortality rate had gone up from an average of 1,000 daily to 4,000 daily. Furthermore, the sequencing data suggests that the prevalence of the highly transmissible Delta variant of Covid-19 among specimens exceeded seventy-five percent in many countries worldwide, including India, China, UK, and Israel. The big question on everyone’s mind is: when will this nightmare end? or will it ever end?

The answer to this question lies in the ability of the virus to mutate into other variants of SARS-CoV-2, thus breaking a person’s hard-won immunity. So future waves are on the cards. Due to the continuous new emerging mutations in the structural proteins or, more specifically, in the spike protein of the virus, it is mandatory to either develop a new vaccine or test the existing vaccines against the emerging variant of the virus. So, do scientists need to develop a new vaccine every time a new variant emerges? Or do they need to develop a vaccine that will work against all the variants? Whatever scientists decide to do, vaccine development is not a simple process. To accommodate the steps of vaccine production, several factors play an important role, such as the difficulty in manufacturing preparations of sufficient titer, increased cost per dose, and current requirements for multiple immunizations. Therefore, developing a new or modified vaccine for the upcoming variants of coronavirus must take place several months in advance.=

To overcome these limitations, Premas Biotech, India, has used its novel platform D-Crypt™, to produce VLPs against UK, SA, and Wuhan variants of SARS-CoV-2, which highlights the potential of D-Crypt™ technology (Figure 1-2). D-Crypt™ platform is an engineered protease deficient strain of S.cerevisiae.

Genetic variants can be easily integrated into expression vectors to modify recombinant proteins rapidly. These innovations make it possible to produce VLPs in just 1-2 months, ensuring their functionality at a larger scale. Therefore, it has added advantages for making a VLP vaccine against new emerging variants of SARS-CoV-2 in shorter times. This system allows the drug product’s scalability, cost-effectiveness and facilitates rapid regulatory approval and straightforward technology transfer to Premas partners.

The design and manufacture of the PRAK-03202, VLP based vaccine candidate against the Wuhan variant of SARS-CoV-2, represent a plug and play process in which three-target antigen (S, E, and M) sequences are inserted into a highly characterized D-Crypt™ platform (Premas Biotech). Although it is worldwide noted that variants of SARS-CoV-2 have mutations in spike protein, only a few mutations have been observed in other structural proteins (envelope and membrane proteins) of SARS-CoV-2. Therefore, the inclusion of M and E antigens would help provide a broader immune response against the variants of SARS-CoV-2. It would also help us tackle the problem of arising mutations in the spike protein of emerging SARS-CoV-2 variants. Furthermore, we at Premas Biotech have shown that immunization using PRAK-03202 increased IgG titers or humoral response (Figure 3A). We next observe the proliferation potential of B cells by flow cytometry at lower doses (1 and 2.5mg). Results showed an increase in proliferation potential of B cells in the group vaccinated with one and 2.5mg of doses compared with the 5 and 10mg doses (Figure 3B). This proliferation potential was based on Ki-67 protein, a cellular marker for proliferation, and is present in the cell during all active phases of the cell cycle (G1, S, G2, and mitosis) but absent from resting cells (G0). These results suggest that the PRAK-03202 can induce both humoral and cell-mediated responses.

In addition to this, we have formed Oravax Medical lnc., a joint venture focused on developing oral VLP based COVID-19 vaccines that target three structural proteins. The joint venture company, Oravax, has combined India-based Premas Biotech’s D-Crypt™ platform with Israel-based Oramed’s POD™ oral delivery technology. It is noteworthy that the oral administration of the vaccine may enable easier distribution and large-scale inoculation without needing an injection.

Figure 3: Humoral and cellular immune responses in immunized rats

A) Five groups of rats were immunized with indicated doses of PRAK-03202 (VLP) either with or without AH (n = 5/group). PRAK-03202 specific IgG antibody titers were analyzed by ELISA in the serum samples. Bars are the mean titer with standard deviation indicated.

B) Bar chart depicting proliferation potential in B lymphocytes in mice vaccinated at 1, 2.5, 5 and 10μg doses of PRAK-03202 (VLP).

Premas Biotech is an Independent Research Organization has the mandate to perform both in-house research activities and contract research activities (CRO). The facility is housed in over 20,000 sq. ft building dedicated to research & development activities and pilot-scale production for research purposes. We have successfully manufactured GMP lots for various drug substances. which are under clinical trials in USA and EMA regions such as Oral insulin (Phase 3), Key molecule to reduce obesity (phase 1 successful data carried out in Israel for the first time in the world). We have also developed a process for ultra-rapid development and scaled-up production of a key enzyme as a Covid-19 diagnostic in 22 days from concept to millions of tests.

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Reference (Feb-22-E1)

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