cGMP production and analysis of BG505 SOSIP.664, an extensively glycosylated, trimeric HIV-1 envelope glycoprotein vaccine candidate

We describe the properties of BG505 SOSIP.664 HIV-1 envelope glycoprotein trimersproduced under current Good Manufacturing Practice(cGMP) conditions. These proteins are the first of a new generation of native-like trimersthat are the basis for many structure-guided immunogen development programs aimed at devising how to induce broadly neutralizing antibodies (bNAbs) to HIV-1 by vaccination. The successful translation of this prototype demonstrates the feasibility of producing similar immunogens on an appropriate scale and of an acceptable quality for Phase I experimental medicine clinical trials. BG505 SOSIP.664 trimersare extensively glycosylated, contain numerous disulfide bondsand require proteolytic cleavage, all properties that pose a substantial challenge to cGMP production. Our strategy involved creating a stable CHO cell line that was adapted to serum-free culture conditions to produce envelope glycoproteins. The trimerswere then purified by chromatographic methods using a 2G12 bNAb affinity column and size-exclusion chromatography. The chosen procedures allowed any adventitious viruses to be cleared from the final product to the required extent of >12 log10 . The final cGMP production run yielded 3.52 grams (peptidic mass) of fully purified trimers(Drug Substance) from a 200 L bioreactor, a notable yield for such a complex glycoprotein. The purified trimerswere fully native-like as judged by negative-stainelectron microscopy, and were stable over a multi-month period at room temperature or below and for at least one week at 50°C. Their antigenicity, disulfide bondpatterns and glycan composition were consistent with trimersproduced on a research laboratory scale. The methods reported here should pave the way for the cGMP production of other native-like Env glycoprotein trimersof various designs and genotypes. This article is protected by copyright. All rights reserved