Glycosylation Benchmark Profile for HIV-1 Envelope Glycoprotein Production Based on Eleven Env Trimers

• DOI: 10.1128/jvi.02428-16
• Publication Date: 2017-02-16T01:50:10Z
• Authors: Eden P. Go; Haitao Ding; Shijian Zhang; Rajesh P. Ringe; Nathan Nicely; David Hua; Robert T. Steinbock; Michael Golabek; James Alin; S. Munir Alam; Albert Cupo; Barton F. Haynes; John C. Kappes; John P. Moore; Joseph G. Sodroski; Heather Desaire
• Keywords: N-linked glycosylation

ABSTRACT HIV-1 envelope glycoprotein (Env) glycosylation is important because individual glycans are components of multiple broadly neutralizing antibody epitopes, while shielding other sites that might otherwise be immunogenic. The on Env influenced by a variety factors, including the genotype protein, cell line used for its expression, and details construct design. Here, we mass spectrometry (MS)-based approach to map complete profile at every site in trimers, accomplishing two goals. (i) We determined which contain conserved glycan profiles across many trimeric Envs. (ii) identified variables impact Env's with divergent glycosylation. Over half gp120 11 different Envs have profile, indicating native consensus does indeed exist among trimers. showed some soluble gp120s gp140s exhibit highly compared Env. also assessed several glycosylation: truncating full-length Env; producing Env, instead more virologically relevant T lymphocytes, CHO cells; purifying chromatographic platforms, nickel-nitrilotriacetic acid (Ni-NTA), 2G12, PGT151 affinity. This report provides first should serve as useful benchmark vaccine developers. defines where may impacted when trimers truncated or produced cells. IMPORTANCE A protective will likely include recombinant version viral (Env). glycosylated, yet developers lacked guidance how assess whether their immunogens optimal following questions still unanswered. What “target” goal generate natively glycosylated protein? exert greatest influence glycosylation? numerous not deviate investigated changing sequence, design, purification method, producer type. data presented here give “glycosylation target” immunogens, they show protein production can