Sameer Kumar Malladi, Unnatiben Rajeshbhai Patel, Raju S Rajmani, Randhir Singh, Suman Pandey, Sahil Kumar, Sara Khaleeq, Petrus Jansen van Vuren, Shane Riddell, Sarah Goldie, Savitha Gayathri, Debajyoti Chakraborty, Parismita Kalita, Ishika Pramanick, Nupur Agarwal, Poorvi Reddy, Nidhi Girish, Aditya Upadhyaya, Mohammad Suhail Khan, Kawkab Kanjo, Madhuraj Bhat, Shailendra Mani, Sankar Bhattacharyya, Samreen Siddiqui, Akansha Tyagi, Sujeet Jha, Rajesh Pandey, Shashank Tripathi, Somnath Dutta, Alexander J McAuley, Nagendrakumar Balasubramanian Singanallur, Seshadri S Vasan, Rajesh P Ringe, Raghavan Varadarajan
The receptor binding domain (RBD) of SARS-CoV-2 is the primary target of neutralizing antibodies. We designed a trimeric, highly thermotolerant glycan engineered RBD by fusion to a heterologous, poorly immunogenic disulfide linked trimerization domain derived from cartilage matrix protein. The protein expressed at a yield of ∼80-100 mg/L in transiently transfected Expi293 cells, as well as CHO and HEK293 stable cell lines and formed homogeneous disulfide-linked trimers. When lyophilized, these possessed remarkable functional stability to transient thermal stress of up to 100 °C and were stable to long-term storage of over 4 weeks at 37 °C unlike an alternative RBD-trimer with a different trimerization domain...
July 14, 2021: ACS Infectious Diseases