Project Title: Optimization of HIV glycoproteins as vaccine candidates
Research overview. The development of prophylactic vaccines against HIV is of paramount importance in the global drive to halt the spread of the virus. Most new vaccine candidates are based on (or contain) versions of the envelope glycoprotein (Env), the target of the desired broadly neutralizing antibodies. Even after the initial testing of a vaccine candidate, there are hurdles associated with production yield, purification strategy and in vitro stability that may hinder its development as a therapeutic compound. The Baerga Lab, in collaboration with Dr. José Lasalde’s lab, aims to optimize the vaccine development pipeline. In a partnership between UPR, a biotechnology start-up (CDI laboratories) and the advisory input of local biopharmaceutical partners (Amgen Manufacturing, Eli Lilly), state-of-the-art analytical tools are used to optimize the production of trimeric gp145, a promising vaccine candidate discovered and partnered between the Henry M. Jackson Foundation for Military Science and the NIH Vaccine Research Program. It has been shown by the Baerga lab that the yields of trimeric gp145 expressed in CHO cells, are consistently low (3-4 mg/L). A suite of analytical tools to quicly assess glycan composition, aggregation, binding activity and protein degradation is used in the process of clone selection and optimization of protein expression conditions. The PR-CLIMB participants will participate in continued efforts involving this particular vaccine candidate by applying a MALDI-TOF MS method for the measurement of glycan composition of trimeric gp145 and evaluating the effect of modifying the media composition on glycan distribution to boost production yields and quality en route to clinical trials.
Skills/Techniques: PR-CLIMB participants will learn critical biochemical techniques for protein production, purification, and analysis.
Skills/Techniques: PR-CLIMB participants will learn critical biochemical techniques for protein production, purification, and analysis.