Translating Innovation to Clinic: End-to-End Bioprocess Development and cGMP Manufacturing of N332-GT5 HIV Vaccine Candidate for First-In-Human Trials (HVTN144).

Authors

Sammaiah Pallerla, Shaunak D Uplekar, Ferenc Boldog, James C Paulson, Sabyasachi Baboo, John R Yates, Wen-Hsin Lee, Gabriel Ozorowski, Joel D Allen, Max Crispin, Chris Cottrell, Andrew B Ward, Varsha Sitaraman, Tim Broderick, Amber Costakes, Nikolette McCombs, Derek Ryan, Leslie Wolfe, Daniel Craig, Kristen Syvertsen, Albert E Price, Jon M Steichen, William R Schief

Published in

Biotechnology and bioengineering. Oct 21, 2026. Epub Oct 21, 2026.

Abstract

The successful translation of rationally designed HIV-1 immunogens into effective vaccines requires manufacturing platforms that maintain structural conformity while meeting clinical-grade quality standards. We developed and scaled up a robust, cGMP-compliant process for N332-GT5 gp140, a germline-targeting envelope trimer designed to initiate broadly neutralizing antibody responses, which is now undergoing first-in-human evaluation in HVTN144. Starting with a stable CHO cell line developed using Leap-In® transposon technology, we established a production clone exhibiting high-titer expression (> 200 mg/L) and genetic stability through 60 population doublings. The manufacturing process scaled up efficiently from Ambr® 250 miniature bioreactors to 200-L single-use systems, delivering consistent product quality across multiple cGMP batches. A streamlined three-step purification strategy-affinity capture, multimodal polishing, and viral clearance -yielded > 99% trimeric purity while preserving quaternary structure and native-like antigenicity. Orthogonal LC-MS analyses confirmed site-specific glycan occupancy matching design specifications, while robust viral clearance exceeded 18-log and 11-log reductions for model retroviruses. Clinical material manufactured through this platform has been successfully administered in clinical trial HVTN144. This work establishes a scalable, reproducible manufacturing paradigm for structurally complex HIV-1 envelope immunogens, advancing the field toward rational vaccine design based on germline-targeting principles.

PMID:
42904530
Bibliographic data and abstract were imported from PubMed on 23 Oct 2026.

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